Antigen binding proteins

ABSTRACT

This application discloses antigen binding proteins that bind Lymphocyte Activation Gene 3 (LAG-3), and more particularly to antigen binding proteins that cause depletion of LAG-3+ activated T cells.

FIELD OF THE INVENTION

The present invention is directed to antigen binding proteins,particularly antibodies that bind Lymphocyte Activation Gene (LAG-3) andcause depletion of activated T cells expressing LAG-3, polynucleotidesencoding such antigen binding proteins, pharmaceutical compositionscontaining said antigen binding proteins, and to the use of said antigenbinding proteins in the treatment and/or prevention of diseasesassociated with the involvement of pathogenic T cells.

BACKGROUND TO THE INVENTION

Lymphocyte Activation Gene-3 (LAG-3) is a negative co-stimulatoryreceptor that modulates T cell homeostasis, proliferation and activation(Sierro S et al; Expert Opin. Ther. Targets (2010) 15: 91-101). Animmunoglobulin superfamily member, LAG-3 is a CD4-like protein which,like CD4, binds to MHC class II molecules, but with two-fold higheraffinity and at a distinct site from CD4 (Huard B et al., (1997) ProcNatl Acad Sci USA 94: 5744-9). In addition to exerting very distinctfunctions, CD4 is a positive co-stimulatory molecule, the two receptorsare also differentially regulated. CD4 is constitutively expressed onthe surface of all mature CD4+ T cells, with only a small fractionresiding intracellularly, whereas a large proportion of LAG-3 moleculesare retained in the cell close to the microtubule-organizing centre, andonly induced following antigen specific T cell activation (Woo S R etal., (2010) Eur J Immunol 40: 1768-77). The role of LAG-3 as a negativeregulator of T cell responses is based on studies with LAG-3 knockoutmice and use of blocking anti-LAG-3 antibodies in model in vitro and invivo systems (Sierro S et al., Expert Opin. Ther. Targets (2010) 15:91-101; Hannier S et al (1998), J Immunol 161: 4058-65; Macon-Lemaitre Let al (2005), Immunology 115: 170-8; Workman C J et al (2003), Eur JImmunol 33:970-9).

At the cell surface, LAG-3 is expressed as a dimer, which is requiredfor formation of stable MHC class II binding sites (Huard B et al.(1997) Proc Natl Acad Sci USA 94: 5744-9). LAG-3, in soluble form, alsooccurs in serum of healthy donors and patients with tuberculosis andcancer (Lienhardt C et al. (2002), Eur J Immunol 32: 1605-13; Triebel Fet al (2006). Cancer Lett 235: 147-53), and this form may correlate withthe number of LAG-3+ T cells (Siawaya J et al. (2008). J of Infection56: 340-7). The key attribute of LAG-3 as a target antigen for anenhanced lymphocyte depletion agent is its relatively selectiveexpression profile when compared with other agents currently in theclinic, i.e. Campath (T/B cells), Amevive (most CD45RO+ T-cells) orRituxan (B-cells). Few molecules have been identified as sustainedmarkers of in vivo T cell activation in humans. These include LAG-3,OX40, MHC class II, CD69, CD38, ICOS and CD40L. However, apart fromLAG-3 and OX40 the majority of these molecules are also constitutivelyexpressed on human natural T regs or on other cell types. LAG-3 isexpressed on a small proportion of T-cells in healthy humans (ca. 1-4%),and in a similar proportion of NK cells (Baixeras E et al. (1992), J ExpMed 176: 327-37; Huard B et al (1994), Immunogenetics 39: 213-7). Uponactivation with anti-CD3 ca. 30-80% of both CD4+ and CD8+ T cellsexpress LAG-3 within 24 to 48 h; this percentage is increased inpresence of IL2, IL7 and IL12 (Sierro S et al, Expert Opin. Ther.Targets (2010) 15: 91-101; Bruniquel D et al (1998), Immunogenetics 48:116-24). Following antigen-specific stimulation with recall antigen(i.e. CMV or Tetanus toxoid) the majority of activated T cells areLAG-3+. In addition, in humans, LAG-3 is expressed on a sub-population(1-10%) of CD4+ CD25+ FoxP3+ T regs in healthy human blood. Thispopulation appears to be functionally suppressive in vitro by cellcontact and IL10 dependent mechanisms and therefore may represent apopulation of recently activated natural or induced T regs [CamisaschiC, Casati C, Rini F et al. (2010). LAG-3 expression defines a subset ofCD4+CD25highFox3P+regulatory T cells that are expanded at tumour sites.J. Immunol. 184: 6545-51). LAG-3 has been detected on other cell typesof hematopoietic lineage, such as plasmacytoid dendritic cells, B-cells,and NKT-cells, but only in the mouse, and mostly following activation(Sierro S, Romero P & Speiser D; Expert Opin. Ther. Targets (2010) 15:91-101).

Depletion of LAG-3+ T cells may be used to treat or prevent T celldriven immuno-inflammatory disorders. In auto-immune diseases where themajority of auto-reactive cells are chronically activated by selfantigens at the disease site and/or re-circulate in the periphery, ashort course of a depleting antigen binding protein may selectivelydeplete this auto-immune T cell repertoire providing long termremission. The precedence for this approach has been demonstrated withthe pan-lymphocyte depleting antibody Campath, in which a single 12 mginjection reduced the rate of relapse by 74% compared to standardtreatment in a multiple sclerosis trial (The CAMMS223 TrialInvestigators (2008), N Engl J Med. 359:1786-801). Due to the moreselective expression of LAG-3 compared with CD52, the target forCampath, the impact on the naïve and resting memory T cell and natural Tregs repertoire should be reduced. This is expected to lead to animproved therapeutic index, maintaining efficacy, but with reduced riskof infection and malignancy as well as onset of auto-immunity associatedwith Campath. Additionally, in a baboon tuberculin skin challenge model,the LAG-3 targeting chimeric antibody IMP731 mediated depletion ofLAG-3+ T-cells, both in the periphery and at the skin challenge site,resulting in a reduction in the tuberculin skin challenge response(Poirier N et al. (2011), Clin Exp Immunol 164: 265-74). In a furtherstudy, a LAG-3 polyclonal antibody depleted LAG-3+ infiltrating T-cellsfrom a rat cardiac allograft and prolonged the survival of these grafts(Haudebourg T et al. (2007), Transplantation 84: 1500-1506).

There exists a need in the art for antigen binding proteins,particularly humanised antibodies, that bind LAG-3 and cause deletion ofLAG-3+ activated T cells, and which may have use in the treatment ofauto-immune diseases, such as psoriasis, Crohn's disease, rheumatoidarthritis, primary biliary cirrhosis, SLE, Sjögren's syndrome, multiplesclerosis, ulcerative colitis and autoimmune hepatitis; and cancer.

SUMMARY OF THE INVENTION

The present invention is broadly directed to antigen binding proteins,such as humanised antibodies, which bind Lymphocyte Activation Gene 3(LAG-3) and which may be able to cause depletion of LAG-3+ activated Tcells. More specifically, antigen binding protein of the presentinvention may comprise CDRL1 of SEQ ID No. 5, wherein position 27E isproline.

Antigen binding proteins described herein may have use in the treatmentor prevention of diseases associated with the involvement of pathogenicT cells, for example auto-immune diseases, such as psoriasis, Crohn'sdisease, rheumatoid arthritis, primary biliary cirrhosis, SLE, Sjögren'ssyndrome, multiple sclerosis, ulcerative colitis and autoimmunehepatitis. Accordingly, the invention is further directed topharmaceutical compositions comprising an antigen binding protein, andoptionally one or more pharmaceutically acceptable excipients and/orcarriers.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A shows a graph of three antibodies (H5L7BW, H5L7 and IMP731)binding to LAG-3 expressing EL4 cells as measured by FACS analysis,using Synagis, a monoclonal antibody against an unrelated target, asnegative control.

FIG. 1B shows a graph of three antibodies (H5L7BW, H5L7 and IMP731)binding to activated human CD3+ T cells as measured by FACS analysis,using Synagis as negative control.

FIG. 2 shows FACS Plots illustrating the gating strategy used toidentify LAG-3 positive antigen specific T cells.

FIG. 3A shows a graph of the quantification of human CD4⁺ LAG-3⁺ andCD8⁺ LAG-3⁺ T cells 24 hours after co-administration of 1×10⁷ activatedhuman PBMCs and 5 mg/kg Control antibody, H5L7BW or MabCampath (Donor A:Control n=2; H5L7BW n=2, MabCampath n=1; Donor B: Control n=2; H5L7BWn=3, MabCampath n=2).

FIG. 3B shows a graph of the quantification of total CD4⁺ and CD8⁺ Tcells (*p<0.001) 24 hours after co-administration of 1×10⁷ activatedhuman PBMCs and 5 mg/kg Control antibody, H5L7BW or MabCampath (Donor A:Control n=2; H5L7BW n=2, MabCampath n=1; Donor B: Control n=2; H5L7BWn=3, MabCampath n=2).

FIG. 4A shows a graph of the quantification of human CD4⁺ LAG-3⁺ andCD8⁺ LAG-3⁺ T cells 5 hours after co-administration of 1×10⁷ activatedhuman PBMCs and 5 mg/kg Control antibody, H5L7BW or H5L7 (n=3 pergroup), which demonstrates the effect of H5L7BW and its non-ADCCenhanced variant H5L7 on activated human PBMCs co-administeredintra-peritoneally and retrieved from the peritoneal cavity 5 hourspost-injection.

FIG. 4B shows a graph of the quantification of total CD4⁺ and CD8⁺ Tcells (*p<0.001) 5 hours after co-administration of 1×10⁷ activatedhuman PBMCs and 5 mg/kg Control antibody, H5L7BW or H5L7 (n=3 pergroup), which demonstrates the effect of H5L7BW and its non-ADCCenhanced variant H5L7 on activated human PBMCs co-administeredintra-peritoneally and retrieved from the peritoneal cavity 5 hourspost-injection.

FIG. 5A shows a graph of the quantification of human CD4⁺ LAG-3⁺ andCD8⁺ LAG-3⁺ T cells 5 hours after i.p. injection of 5×10⁶ (n=1 pergroup) or 1×10⁷ O/N (n=4 per group) activated human PBMCs into SCIDmice, where H5L7BW was administered intravenously 18 hours beforeadministration of human activated PBMCs into the peritoneum of SCIDmice.

FIG. 5B shows a graph of the quantification of total CD4⁺ and CD8⁺ Tcells 5 hours after i.p. injection of 5×10⁶ (n=1 per group) or 1×10⁷ O/N(n=4 per group) activated human PBMCs into SCID mice, where 5 mg/kgH5L7BW or control antibody was injected intravenously 18 hours beforeadministration of activated human PBMCs. (*p<0.001, #p=0.0052)

FIG. 6A shows a graph of the quantification of human CD4⁺ LAG-3⁺ andCD8⁺LAG-3⁺ T cells 5 hours after i.p. injection of 1×10⁷ (n=4 per group)activated human PBMCs into SCID mice, where 5 mg/kg H5L7BW, H5L7, IMP731or control antibody were injected intravenously 18 hours beforeadministration of human PBMCs.

FIG. 6B shows a graph of the quantification of total CD4⁺ and CD8⁺ Tcells 5 hours after i.p. injection of 1×10⁷ n=4 per group) activatedhuman PBMCs into SCID mice, where 5 mg/kg LAG-3 depleting antibodies orcontrol antibody were injected intravenously 18 hours pre-injection ofactivated human PBMCs. (*p<0.001)

DETAILED DESCRIPTION OF THE INVENTION

The present invention is broadly directed to antigen binding proteinsthat bind Lymphocyte Activation Gene 3 (LAG-3), and more particularly toantigen binding proteins that may cause depletion of LAG-3+ activated Tcells.

The term “antigen binding protein” as used herein refers to antibodiesand fragments thereof which are capable of binding to LAG-3. Unlessotherwise specified, the term “LAG-3” as used herein refers toLymphocyte Activation Gene 3, expressed as a dimer on the surface of,for example, activated T cells, NK cells and B cells. The term “sLAG-3”as used herein refers to a soluble form of LAG-3 found, for example, inserum. Unless otherwise specified, references herein to “sLAG-3” and“LAG-3” are to human polypeptides.

In one aspect, the present invention provides antigen binding proteinsthat are capable of binding LAG-3 and which comprise CDRL1 of SEQ ID No.5, wherein position 27E is proline.

In a further aspect, the present invention provides antigen bindingproteins, which comprises CDRL1 of SEQ ID NO. 1.

In a further aspect, the present invention provides antigen bindingproteins that in addition to comprising CDRL1 of SEQ ID NO.5, alsocomprise CDRL2 and/or CDRL3 from SEQ ID NO. 5, or a CDR variant thereof.

In a further aspect, the present invention provides antigen bindingproteins comprising CDRL2 of SEQ ID NO. 2 and/or CDRL3 of SEQ ID NO. 3or a CDR variant thereof.

In a further aspect, the present invention provides antigen bindingproteins comprising one or more of CDRH1, CDRH2 and CDRH3, or a CDRvariant thereof, from SEQ ID NO 10.

In a further aspect, the present invention provides antigen bindingproteins comprising one or more CDRs, or a CDR variant thereof, selectedfrom the group comprising CDRH1 of SEQ ID NO. 6, CDRH2 of SEQ ID NO. 7and CDRH3 of SEQ ID NO. 8.

In a further aspect, the present invention provides antigen bindingproteins comprising the following CDRs:

CDRL1: SEQ ID NO. 1

CDRL2: SEQ ID NO. 2

CDRL3: SEQ ID NO. 3

CDRH1: SEQ ID NO. 6

CDRH2: SEQ ID NO. 7

CDRH3: SEQ ID NO. 8

The term “CDR” as used herein, refers to the complementarity determiningregion amino acid sequences of an antigen binding protein. These are thehypervariable regions of immunoglobulin heavy and light chains. Thereare three heavy chain and three light chain CDRs (or CDR regions) in thevariable portion of an immunoglobulin.

It will be apparent to those skilled in the art that there are variousnumbering conventions for CDR sequences; Chothia (Chothia et al. (1989)Nature 342: 877-883), Kabat (Kabat et al., Sequences of Proteins ofImmunological Interest, 4th Ed., U.S. Department of Health and HumanServices, National Institutes of Health (1987)), AbM (University ofBath) and Contact (University College London). The minimum overlappingregion using at least two of the Kabat, Chothia, AbM and contact methodscan be determined to provide the “minimum binding unit”. The minimumbinding unit may be a sub-portion of a CDR. The structure and proteinfolding of the antibody may mean that other residues are considered partof the CDR sequence and would be understood to be so by a skilledperson.

Unless otherwise stated and/or in absence of a specifically identifiedsequence, references herein to “CDR”, “CDRL1”, “CDRL2”, “CDRL3”,“CDRH1”, “CDRH2”, “CDRH3” refer to amino acid sequences numberedaccording to any of the known conventions identified in Table 1.

Table 1 below represents one definition using each numbering conventionfor each CDR or binding unit. It should be noted that some of the CDRdefinitions may vary depending on the individual publication used.

TABLE 1 Kabat Chothia AbM Contact Minimum CDR CDR CDR CDR binding unitH1 31-35/ 26-32/ 26-35/ 30-35/ 31-32 35A/35B 33/34 35A/35B 35A/35B H250-65 52-56 50-58 47-58 52-56 H3  95-102  95-102  95-102  93-101  95-101L1 24-34 24-34 24-34 30-36 30-34 L2 50-56 50-56 50-56 46-55 50-55 L389-97 89-97 89-97 89-96 89-96

The term “CDR variant” as used herein, refers to a CDR that has beenmodified by at least one, for example 1, 2 or 3, amino acidsubstitution(s), deletion(s) or addition(s), wherein the modifiedantigen binding protein comprising the CDR variant substantially retainsthe biological characteristics of the antigen binding proteinpre-modification. It will be appreciated that each CDR that can bemodified may be modified alone or in combination with another CDR. Inone aspect, the modification is a substitution, particularly aconservative substitution, for example as shown in Table 2.

TABLE 2 Side chain Members Hydrophobic Met, Ala, Val, Leu, Ile Neutralhydrophilic Cys, Ser, Thr Acidic Asp, Glu Basic Asn, Gln, His, Lys, ArgResidues that influence chain orientation Gly, Pro Aromatic Trp, Tyr,Phe

For example, in a variant CDR, the amino acid residues of the minimumbinding unit may remain the same, but the flanking residues thatcomprise the CDR as part of the Kabat or Chothia definition(s) may besubstituted with a conservative amino acid residue.

Such antigen binding proteins comprising modified CDRs or minimumbinding units as described above may be referred to herein as“functional CDR variants” or “functional binding unit variants”.

Antigen binding proteins of the present invention may be capable ofbinding sLAG-3. In one aspect, the equilibrium dissociation constant(KD) of the antigen binding protein-sLAG-3 interaction is 10 nM or less,such as 1 nM or less, for example between 1 pM and 300, 400, 500 pM orbetween 500 pM and 1 nM. A skilled person will appreciate that thesmaller the KD numerical value, the stronger the binding. The reciprocalof KD (i.e. 1/KD) is the equilibrium association constant (KA) havingunits M⁻¹. A skilled person will appreciate that the larger the KAnumerical value, the stronger the binding.

In one aspect, the present invention provides antigen binding proteinsthat are capable of binding recombinant LAG-3 with a KD of less than 1nM, for example between 1 μM and 300 μM, when determined by Biacore™surface Plasmon resonance analysis using recombinant human, orcynomolgus macaque LAG-3 extracellular domains (ECDs) of SEQ ID NOs:51and 52, respectively.

Furthermore, antigen binding proteins of the present invention may alsobe capable of binding LAG-3 expressed on, for example, EL4 cells oractivated human CD3+ T cells.

Antigen binding proteins of the present invention may also be capable ofdepleting LAG-3+ activated T cells, in particular, CD4+LAG-3+ andCD8+LAG-3+ T cells. Depletion of LAG-3+ T cells may occur by, forexample, antibody dependent cell mediated cytotoxic activity (ADCC)and/or complement-dependent cytotoxic activity (CDC).

In one aspect, the present invention provides antigen binding proteinsthat are capable of causing greater than 40% depletion of antigenspecific CD4 and/or CD8 LAG-3⁺ human T cells by ADCC in an in-vitroassay using primary human T cells.

In a further aspect, the present invention provides antigen bindingproteins that, at a concentration of 0.1 μg/mL, are capable of causinggreater than 50% depletion in an in vitro ADCC assay usingeuropium-labelled LAG-3 expressing EL4 cells as target cells and humanPBMCs as effector cells, wherein the effector:target ratio is no greaterthan 50:1 and the assay is run for a period of 2 hours. % cell lysis iscalculated based on europium release from LAG-3 expressing EL4 cells.

The interaction between the constant region of an antigen bindingprotein and various Fc receptors (FcR) including FcγRI (CD64), FcγRII(CD32) and FcγRIII (CD16) is believed to mediate the effector functions,such as ADCC and CDC, of the antigen binding protein. Significantbiological effects can be a consequence of effector functionality.Usually, the ability to mediate effector function requires binding ofthe antigen binding protein to an antigen and not all antigen bindingproteins will mediate every effector function.

Effector function can be measured in a number of ways including forexample via binding of the FcγRIII to Natural Killer cells or via FcγRIto monocytes/macrophages to measure for ADCC effector function. Forexample an antigen binding protein of the present invention can beassessed for ADCC effector function in a Natural Killer cell assay.Examples of such assays can be found in Shields et al, 2001 The Journalof Biological Chemistry, Vol. 276, p 6591-6604; Chappel et al, 1993 TheJournal of Biological Chemistry, Vol 268, p 25124-25131; Lazar et al,2006 PNAS, 103; 4005-4010.

Examples of assays to determine CDC function include that described in1995 J Imm Meth 184:29-38.

In one aspect of the present invention, the antigen binding protein isan antibody.

The term “antibody” as used herein refers to molecules with animmunoglobulin-like domain and includes monoclonal (for example IgG,IgM, IgA, IgD or IgE), recombinant, polyclonal, chimeric, humanised,bispecific and heteroconjugate antibodies; a single variable domain(e.g., VH, VHH, VL), a domain antibody (dAb®), antigen bindingfragments, immunologically effective fragments, Fab, F(ab′)₂, Fv,disulphide linked Fv, single chain Fv, closed conformation multispecificantibody, disulphide-linked scFv, diabodies, TANDABS™, etc. and modifiedversions of any of the foregoing (for a summary of alternative“antibody” formats see Holliger and Hudson, Nature Biotechnology, 2005,Vol 23, No. 9, 1126-1136). Alternative antibody formats includealternative scaffolds in which the one or more CDRs of any molecules inaccordance with the disclosure can be arranged onto a suitablenon-immunoglobulin protein scaffold or skeleton, such as an affibody, aSpA scaffold, an LDL receptor class A domain, an avimer (see, e.g., U.S.Patent Application Publication Nos. 2005/0053973, 2005/0089932,2005/0164301) or an EGF domain.

In a further aspect, the antigen binding protein is a humanisedantibody.

A “humanised antibody” refers to a type of engineered antibody havingits CDRs derived from a non-human donor immunoglobulin, the remainingimmunoglobulin-derived parts of the molecule being derived from one (ormore) human immunoglobulin(s). In addition, framework support residuesmay be altered to preserve binding affinity (see, e.g., Queen et al.,Proc. Natl. Acad Sci USA, 86:10029-10032 (1989), Hodgson et al.,Bio/Technology, 9:421 (1991)). A suitable human acceptor antibody may beone selected from a conventional database, e.g., the KABAT® database,Los Alamos database, and Swiss Protein database, by homology to thenucleotide and amino acid sequences of the donor antibody. A humanantibody characterized by a homology to the framework regions of thedonor antibody (on an amino acid basis) may be suitable to provide aheavy chain constant region and/or a heavy chain variable frameworkregion for insertion of the donor CDRs. A suitable acceptor antibodycapable of donating light chain constant or variable framework regionsmay be selected in a similar manner. It should be noted that theacceptor antibody heavy and light chains are not required to originatefrom the same acceptor antibody. The prior art describes several ways ofproducing such humanised antibodies—see for example EP-A-0239400 andEP-A-054951.

In yet a further aspect, the humanised antibody has a human antibodyconstant region that is an IgG1, for example, the heavy chain constantregion of SEQ ID No. 46.

It will be understood that the present invention further provideshumanised antibodies which comprise a) a light chain sequence of SEQ IDNO. 5 or a light chain sequence with at least 75%, 80%, 85%, 90%, 95%,96%, 97%, 98% or 99% identity to any of SEQ ID NO. 5 and b) a heavychain sequence of SEQ ID NO. 10 or a heavy chain sequence with at least75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity to any of SEQ IDNO. 10.

It will be understood that the present invention further provideshumanised antibodies which comprise a) a light chain sequence of SEQ IDNO. 35 or a light chain sequence with at least 75%, 80%, 85%, 90%, 95%,96%, 97%, 98% or 99% identity to any of SEQ ID NO. 35 and b) a heavychain sequence of SEQ ID NO. 36 or a heavy chain sequence with at least75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity to any of SEQ IDNO. 36.

In a further aspect, the present invention provides antigen bindingproteins comprising CDRL1-L3 and CDRH1-H3 of SEQ ID NO: 35 and 36,respectively. For nucleotide and amino acid sequences, the term“identical” or “identity” indicates the degree of identity between twonucleic acid or two amino acid sequences when optimally aligned andcompared with appropriate insertions or deletions.

The percent identity between two sequences is a function of the numberof identical positions shared by the sequences (i.e., % identity=numberof identical positions/total number of positions multiplied by 100),taking into account the number of gaps, and the length of each gap,which need to be introduced for optimal alignment of the two sequences.The comparison of sequences and determination of percent identitybetween two sequences can be accomplished using a mathematicalalgorithm, as described below.

Percent identity between a query nucleic acid sequence and a subjectnucleic acid sequence is the “Identities” value, expressed as apercentage, which is calculated by the BLASTN algorithm when a subjectnucleic acid sequence has 100% query coverage with a query nucleic acidsequence after a pair-wise BLASTN alignment is performed. Such pair-wiseBLASTN alignments between a query nucleic acid sequence and a subjectnucleic acid sequence are performed by using the default settings of theBLASTN algorithm available on the National Center for BiotechnologyInstitute's website with the filter for low complexity regions turnedoff. Importantly, a query nucleic acid sequence may be described by anucleic acid sequence identified in one or more claims herein.

Percent identity between a query amino acid sequence and a subject aminoacid sequence is the “Identities” value, expressed as a percentage,which is calculated by the BLASTP algorithm when a subject amino acidsequence has 100% query coverage with a query amino acid sequence aftera pair-wise BLASTP alignment is performed. Such pair-wise BLASTPalignments between a query amino acid sequence and a subject amino acidsequence are performed by using the default settings of the BLASTPalgorithm available on the National Center for Biotechnology Institute'swebsite with the filter for low complexity regions turned off.Importantly, a query amino acid sequence may be described by an aminoacid sequence identified in one or more claims herein.

In a further aspect, the present invention provides a humanisedantibody, which comprises a) a light chain sequence with at least 90%identity to SEQ ID NO. 5, and b) a heavy chain sequence with at least90% identity to SEQ ID NO. 10.

In a further aspect, the present invention provides a humanisedantibody, which comprises a) a light chain sequence with at least 95%identity to SEQ ID NO. 5, and b) a heavy chain sequence with at least95% identity to SEQ ID NO. 10.

In yet a further aspect, the present invention provides a humanisedantibody, which comprises a) a light chain sequence with at least 97%identity to SEQ ID NO. 5, and b) a heavy chain sequence with at least97% identity to SEQ ID NO. 10.

In yet a further aspect, the present invention provides a humanisedantibody, which comprises a) a light chain sequence of SEQ ID NO. 5, andb) a heavy chain sequence of SEQ ID NO. 10.

Production

The antigen binding proteins, for example antibodies of the presentinvention may be produced by transfection of a host cell with anexpression vector comprising the coding sequence for the antigen bindingprotein of the invention. An expression vector or recombinant plasmid isproduced by placing these coding sequences for the antigen bindingprotein in operative association with conventional regulatory controlsequences capable of controlling the replication and expression in,and/or secretion from, a host cell. Regulatory sequences includepromoter sequences, e.g., CMV promoter, and signal sequences which canbe derived from other known antibodies. Similarly, a second expressionvector can be produced having a DNA sequence which encodes acomplementary antigen binding protein light or heavy chain. In certainembodiments this second expression vector is identical to the firstexcept insofar as the coding sequences and selectable markers areconcerned, so to ensure as far as possible that each polypeptide chainis functionally expressed. Alternatively, the heavy and light chaincoding sequences for the antigen binding protein may reside on a singlevector.

A selected host cell is co-transfected by conventional techniques withboth the first and second vectors (or simply transfected by a singlevector) to create the transfected host cell of the invention comprisingboth the recombinant or synthetic light and heavy chains. Thetransfected cell is then cultured by conventional techniques to producethe engineered antigen binding protein of the invention. The antigenbinding protein which includes the association of both the recombinantheavy chain and/or light chain is screened from culture by appropriateassay, such as ELISA or RIA. Similar conventional techniques may beemployed to construct other antigen binding proteins.

Suitable vectors for the cloning and subcloning steps employed in themethods and construction of the compositions of this invention may beselected by one of skill in the art. For example, the conventional pUCseries of cloning vectors may be used. One vector, pUC19, iscommercially available from supply houses, such as Amersham(Buckinghamshire, United Kingdom) or Pharmacia (Uppsala, Sweden).Additionally, any vector which is capable of replicating readily, has anabundance of cloning sites and selectable genes (e.g., antibioticresistance), and is easily manipulated may be used for cloning. Thus,the selection of the cloning vector is not a limiting factor in thisinvention.

The expression vectors may also be characterized by genes suitable foramplifying expression of the heterologous DNA sequences, e.g., themammalian dihydrofolate reductase gene (DHFR). Other preferable vectorsequences include a poly A signal sequence, such as from bovine growthhormone (BGH) and the betaglobin promoter sequence (betaglopro). Theexpression vectors useful herein may be synthesized by techniques wellknown to those skilled in this art.

The components of such vectors, e.g. replicons, selection genes,enhancers, promoters, signal sequences and the like, may be obtainedfrom commercial or natural sources or synthesized by known proceduresfor use in directing the expression and/or secretion of the product ofthe recombinant DNA in a selected host. Other appropriate expressionvectors of which numerous types are known in the art for mammalian,bacterial, insect, yeast, and fungal expression may also be selected forthis purpose.

The present invention also encompasses a cell line transfected with arecombinant plasmid containing the coding sequences of the antigenbinding proteins of the present invention. Host cells useful for thecloning and other manipulations of these cloning vectors are alsoconventional. However, cells from various strains of E. coli may be usedfor replication of the cloning vectors and other steps in theconstruction of antigen binding proteins of this invention.

Suitable host cells or cell lines for the expression of the antigenbinding proteins of the invention include mammalian cells such as NS0,Sp2/0, CHO (e.g. DG44), COS, HEK, a fibroblast cell (e.g., 3T3), andmyeloma cells, for example it may be expressed in a CHO or a myelomacell. Human cells may be used, thus enabling the molecule to be modifiedwith human glycosylation patterns. Alternatively, other eukaryotic celllines may be employed. The selection of suitable mammalian host cellsand methods for transformation, culture, amplification, screening andproduct production and purification are known in the art. See, e.g.,Sambrook et al., cited above.

Bacterial cells may prove useful as host cells suitable for theexpression of the recombinant Fabs or other embodiments of the presentinvention (see, e.g., Plückthun, A., Immunol. Rev., 130:151-188 (1992)).However, due to the tendency of proteins expressed in bacterial cells tobe in an unfolded or improperly folded form or in a non-glycosylatedform, any recombinant Fab produced in a bacterial cell would have to bescreened for retention of antigen binding ability. If the moleculeexpressed by the bacterial cell was produced in a properly folded form,that bacterial cell would be a desirable host, or in alternativeembodiments the molecule may express in the bacterial host and then besubsequently re-folded. For example, various strains of E. coli used forexpression are well-known as host cells in the field of biotechnology.Various strains of B. subtilis, Streptomyces, other bacilli and the likemay also be employed in this method.

Where desired, strains of yeast cells known to those skilled in the artare also available as host cells, as well as insect cells, e.g.Drosophila and Lepidoptera and viral expression systems. See, e.g.Miller et al., Genetic Engineering, 8:277-298, Plenum Press (1986) andreferences cited therein.

The general methods by which the vectors may be constructed, thetransfection methods required to produce the host cells of theinvention, and culture methods necessary to produce the antigen bindingprotein of the invention from such host cell may all be conventionaltechniques. Typically, the culture method of the present invention is aserum-free culture method, usually by culturing cells serum-free insuspension. Likewise, once produced, the antigen binding proteins of theinvention may be purified from the cell culture contents according tostandard procedures of the art, including ammonium sulfateprecipitation, affinity columns, column chromatography, gelelectrophoresis and the like. Such techniques are within the skill ofthe art and do not limit this invention. For example, preparations ofaltered antibodies are described in WO 99/58679 and WO 96/16990.

Yet another method of expression of the antigen binding proteins mayutilize expression in a transgenic animal, such as described in U.S.Pat. No. 4,873,316. This relates to an expression system using theanimals casein promoter which when transgenically incorporated into amammal permits the female to produce the desired recombinant protein inits milk. In a further aspect of the invention there is provided amethod of producing an antibody of the invention which method comprisesthe step of culturing a host cell transformed or transfected with avector encoding the light and/or heavy chain of the antibody of theinvention and recovering the antibody thereby produced.

In accordance with the present invention there is provided a method ofproducing an anti-LAG-3 antibody of the present invention which binds tohuman LAG-3, which method comprises the steps of;

-   -   (a) providing a first vector encoding a heavy chain of the        antibody;    -   (b) providing a second vector encoding a light chain of the        antibody;    -   (c) transforming a mammalian host cell (e.g. CHO) with said        first and second vectors;    -   (d) culturing the host cell of step (c) under conditions        conducive to the secretion of the antibody from said host cell        into said culture media;    -   (e) recovering the secreted antibody of step (d).

Once expressed by the desired method, the antibody may then be examinedfor in vitro activity by use of an appropriate assay, such as Biacore™surface Plasmon resonance analysis, to assess binding of the antibody toLAG-3. Additionally, other in vitro and in vivo assays may also be usedto determine an antibody's ability to cause depletion of cellsexpressing LAG-3, such as activated human T cell populations.

The skilled person will appreciate that, upon production of an antigenbinding protein such as an antibody, in particular depending on the cellline used and particular amino acid sequence of the antigen bindingprotein, post-translational modifications may occur. For example, thismay include the cleavage of certain leader sequences, the addition ofvarious sugar moieties in various glycosylation and phosphorylationpatterns, deamidation, oxidation, disulfide bond scrambling,isomerisation, C-terminal lysine clipping, and N-terminal glutaminecyclisation. The present invention encompasses the use of antigenbinding proteins which have been subjected to, or have undergone, one ormore post-translational modifications. Thus an “antigen binding protein”or “antibody” of the invention includes an “antigen binding protein” or“antibody”, respectively, as defined earlier which has undergone apost-translational modification such as described herein.

Glycosylation of antibodies at conserved positions in their constantregions is known to have a profound effect on antibody function,particularly effector functioning, see for example, Boyd et al. (1996)Mol. Immunol. 32: 1311-1318. Glycosylation variants of the antigenbinding proteins of the invention wherein one or more carbohydratemoiety is added, substituted, deleted or modified are contemplated.Introduction of an asparagine-X-serine or asparagine-X-threonine motifcreates a potential site for enzymatic attachment of carbohydratemoieties and may therefore be used to manipulate the glycosylation of anantibody. In Raju et al. (2001) Biochemistry 40: 8868-8876 the terminalsialyation of a TNFR-IgG immunoadhesin was increased through a processof regalactosylation and/or resialylation usingbeta-1,4-galactosyltransferace and/or alpha, 2,3 sialyltransferase.Increasing the terminal sialylation is believed to increase thehalf-life of the immunoglobulin. Antibodies, in common with mostglycoproteins, are typically produced as a mixture of glycoforms. Thismixture is particularly apparent when antibodies are produced ineukaryotic, particularly mammalian cells. A variety of methods have beendeveloped to manufacture defined glycoforms, see Zhang et al. (2004)Science 303: 371: Sears et al. (2001) Science 291: 2344; Wacker et al.(2002) Science 298: 1790; Davis et al. (2002) Chem. Rev. 102: 579; Hanget al. (2001) Acc. Chem. Res 34: 727. The antibodies (for example of theIgG isotype, e.g. IgG1) as herein described may comprise a definednumber (e.g. 7 or less, for example 5 or less, such as two or a single)of glycoform(s).

Deamidation is an enzymatic reaction primarily converting asparagine (N)to iso-aspartic acid and aspartic acid (D) at approximately 3:1 ratio.To a much lesser degree, deamidation can occur with glutamine residuesin a similar manner. Deamidation in a CDR results in a change in chargeof the molecule, but typically does not result in a change in antigenbinding, nor does it impact on PK/PD.

Oxidation can occur during production and storage (i.e. in the presenceof oxidizing conditions) and results in a covalent modification of aprotein, induced either directly by reactive oxygen species orindirectly by reaction with secondary by-products of oxidative stress.Oxidation happens primarily with methionine residues, but occasionallycan occur at tryptophan and free cysteine residues.

Disulfide bond scrambling can occur during production and basic storageconditions. Under certain circumstances, disulfide bonds can break orform incorrectly, resulting in unpaired cysteine residues (—SH). Thesefree (unpaired) sulfhydryls (—SH) can promote shuffling.

Isomerization typically occurs during production, purification, andstorage (at acidic pH) and usually occurs when aspartic acid isconverted to isoaspartic acid through a chemical process.

N-terminal glutamine in the heavy chain and/or light chain is likely toform pyroglutamate (pGlu). Most pGlu formation happens in the productionbioreactor, but it can be formed non-enzymatically, depending on pH andtemperature of processing and storage conditions. pGlu formation isconsidered as one of the principal degradation pathways for recombinantmAbs.

C-terminal lysine clipping is an enzymatic reaction catalyzed bycarboxypeptidases, and is commonly observed in recombinant mAbs.Variants of this process include removal of lysine from one or bothheavy chains. Lysine clipping does not appear to impact bioactivity andhas no effect on mAb effector function.

Effector Function Enhancement

The interaction between the constant region of an antigen bindingprotein and various Fc receptors (FcR) including FcγRI (CD64), FcγRII(CD32) and FcγRIII (CD16) is believed to mediate the effector functions,such as ADCC and CDC, of the antigen binding protein.

The term “Effector Function” as used herein is meant to refer to one ormore of Antibody dependant cell mediated cytotoxic activity (ADCC),Complement-dependant cytotoxic activity (CDC) mediated responses,Fc-mediated phagocytosis or antibody dependant cellular phagocytosis(ADCP) and antibody recycling via the FcRn receptor.

The ADCC or CDC properties of antigen binding proteins of the presentinvention may be enhanced in a number of ways.

Human IgG1 constant regions containing specific mutations or alteredglycosylation on residue Asn297 have been shown to enhance binding to Fcreceptors. In some cases these mutations have also been shown to enhanceADCC and CDC (Lazar et al. PNAS 2006, 103; 4005-4010; Shields et al. JBiol Chem 2001, 276; 6591-6604; Nechansky et al. Mol Immunol, 2007, 44;1815-1817).

In one embodiment of the present invention, such mutations are in one ormore of positions selected from 239, 332 and 330 (IgG1), or theequivalent positions in other IgG isotypes. Examples of suitablemutations are S239D and I332E and A330L. In one embodiment, the antigenbinding protein of the invention herein described is mutated atpositions 239 and 332, for example S239D and I332E or in a furtherembodiment it is mutated at three or more positions selected from 239and 332 and 330, for example S239D and I332E and A330L. (EU indexnumbering).

In one embodiment of the present invention, there is provided an antigenbinding protein comprising a chimaeric heavy chain constant region forexample an antigen binding protein comprising a chimaeric heavy chainconstant region with at least one CH2 domain from IgG3 such that theantigen binding protein has enhanced effector function, for examplewherein it has enhanced ADCC or enhanced CDC, or enhanced ADCC and CDCfunctions. In one such embodiment, the antigen binding protein maycomprise one CH2 domain from IgG3 or both CH2 domains may be from IgG3.

Also provided is a method of producing an antigen binding proteinaccording to the invention comprising the steps of:

a) culturing a recombinant host cell comprising an expression vectorcomprising an isolated nucleic acid as described herein wherein theexpression vector comprises a nucleic acid sequence encoding an Fcdomain having both IgG1 and IgG3 Fc domain amino acid residues; and

b) recovering the antigen binding protein.

Such methods for the production of antigen binding proteins can beperformed, for example, using the COMPLEGENT™ technology systemavailable from BioWa, Inc. (Princeton, N.J.) and Kyowa Hakko Kogyo (now,Kyowa Hakko Kirin Co., Ltd.) Co., Ltd. In which a recombinant host cellcomprising an expression vector in which a nucleic acid sequenceencoding a chimeric Fc domain having both IgG1 and IgG3 Fc domain aminoacid residues is expressed to produce an antigen binding protein havingenhanced complement dependent cytotoxicity (CDC) activity that isincreased relative to an otherwise identical antigen binding proteinlacking such a chimeric Fc domain. Aspects of the COMPLEGENT™ technologysystem are described in WO2007011041 and US20070148165 each of which areincorporated herein by reference. In an alternative embodiment CDCactivity may be increased by introducing sequence specific mutationsinto the Fc region of an IgG chain. Those of ordinary skill in the artwill also recognize other appropriate systems.

In an alternative embodiment of the present invention, there is providedan antigen binding protein comprising a heavy chain constant region withan altered glycosylation profile such that the antigen binding proteinhas enhanced effector function. For example, wherein the antigen bindingprotein has enhanced ADCC or enhanced CDC or wherein it has bothenhanced ADCC and CDC effector function. Examples of suitablemethodologies to produce antigen binding proteins with an alteredglycosylation profile are described in WO2003011878, WO2006014679 andEP1229125, all of which can be applied to the antigen binding proteinsof the present invention.

The present invention also provides a method for the production of anantigen binding protein according to the invention comprising the stepsof:

a) culturing a recombinant host cell comprising an expression vectorcomprising the isolated nucleic acid as described herein, wherein theFUT8 gene encoding alpha-1,6-fucosyltransferase has been inactivated inthe recombinant host cell; and

b) recovering the antigen binding protein.

Such methods for the production of antigen binding proteins can beperformed, for example, using the POTELLIGENT™ technology systemavailable from BioWa, Inc. (Princeton, N.J.) in which CHOK1SV cellslacking a functional copy of the FUT8 gene produce monoclonal antibodieshaving enhanced antibody dependent cell mediated cytotoxicity (ADCC)activity that is increased relative to an identical monoclonal antibodyproduced in a cell with a functional FUT8 gene. Aspects of thePOTELLIGENT™ technology system are described in US7214775, US6946292,WO0061739 and WO0231240 all of which are incorporated herein byreference. Those of ordinary skill in the art will also recognize otherappropriate systems.

In a further aspect, the present invention provides non-fucosylatedantibodies. Non-fucosylated antibodies harbour a tri-mannosyl corestructure of complex-type N-glycans of Fc without fucose residue. Theseglycoengineered antibodies that lack core fucose residue from the FcN-glycans may exhibit stronger ADCC than fucosylated equivalents due toenhancement of FcγRIIIa binding capacity.

It will be apparent to those skilled in the art that such modificationsmay not only be used alone but may be used in combination with eachother in order to further enhance effector function.

In one such embodiment of the present invention there is provided anantigen binding protein comprising a heavy chain constant region whichcomprises a mutated and chimaeric heavy chain constant region forexample wherein an antigen binding protein comprising at least one CH2domain from IgG3 and one CH2 domain from IgG1, wherein the IgG1 CH2domain has one or more mutations at positions selected from 239 and 332and 330 (for example the mutations may be selected from S239D and I332Eand A330L) such that the antigen binding protein has enhanced effectorfunction, for example wherein it has one or more of the followingfunctions, enhanced ADCC or enhanced CDC, for example wherein it hasenhanced ADCC and enhanced CDC. In one embodiment the IgG1 CH2 domainhas the mutations S239D and I332E.

In an alternative embodiment of the present invention there is providedan antigen binding protein comprising a chimaeric heavy chain constantregion and which has an altered glycosylation profile. In one suchembodiment the heavy chain constant region comprises at least one CH2domain from IgG3 and one CH2 domain from IgG1 and has an alteredglycosylation profile such that the ratio of fucose to mannose is 0.8:3or less, for example wherein the antigen binding protein isdefucosylated so that said antigen binding protein has an enhancedeffector function in comparison with an equivalent antigen bindingprotein with an immunoglobulin heavy chain constant region lacking saidmutations and altered glycosylation profile, for example wherein it hasone or more of the following functions, enhanced ADCC or enhanced CDC,for example wherein it has enhanced ADCC and enhanced CDC.

In an alternative embodiment the antigen binding protein has at leastone IgG3 CH2 domain and at least one heavy chain constant domain fromIgG1 wherein both IgG CH2 domains are mutated in accordance with thelimitations described herein.

In one aspect of the invention there is provided a method of producingan antigen binding protein according to the invention described hereincomprising the steps of:

a) culturing a recombinant host cell containing an expression vectorcontaining an isolated nucleic acid as described herein, said expressionvector further comprising a Fc nucleic acid sequence encoding a chimericFc domain having both IgG1 and IgG3 Fc domain amino acid residues, andwherein the FUT8 gene encoding alpha-1,6-fucosyltransferase has beeninactivated in the recombinant host cell; and

b) recovering the antigen binding protein.

Such methods for the production of antigen binding proteins can beperformed, for example, using the ACCRETAMAB™ technology systemavailable from BioWa, Inc. (Princeton, N.J.) which combines thePOTELLIGENT™ and COMPLEGENT™ technology systems to produce an antigenbinding protein having both ADCC and CDC enhanced activity that isincreased relative to an otherwise identical monoclonal antibody lackinga chimeric Fc domain and which has fucose on the oligosaccharide

In yet another embodiment of the present invention there is provided anantigen binding protein comprising a mutated and chimeric heavy chainconstant region wherein said antigen binding protein has an alteredglycosylation profile such that the antigen binding protein has enhancedeffector function, for example wherein it has one or more of thefollowing functions, enhanced ADCC or enhanced CDC. In one embodimentthe mutations are selected from positions 239 and 332 and 330, forexample the mutations are selected from S239D and I332E and A330L. In afurther embodiment the heavy chain constant region comprises at leastone CH2 domain from IgG3 and one Ch2 domain from IgG1. In one embodimentthe heavy chain constant region has an altered glycosylation profilesuch that the ratio of fucose to mannose is 0.8:3 or less for examplethe antigen binding protein is defucosylated, so that said antigenbinding protein has an enhanced effector function in comparison with anequivalent non-chimaeric antigen binding protein or with animmunoglobulin heavy chain constant region lacking said mutations andaltered glycosylation profile.

Half-Life Extension

Increased half-life, or half-life extension, can be useful in in vivoapplications of antigen binding proteins, especially antibodies and mostespecially antibody fragments of small size. Such fragments (Fvs,disulphide bonded Fvs, Fabs, scFvs, dAbs) are generally rapidly clearedfrom the body. Antigen binding proteins in accordance with thedisclosure can be adapted or modified to provide increased serumhalf-life in vivo and consequently longer persistence, or residence,times of the functional activity of the antigen binding protein in thebody. Suitably, such modified molecules have a decreased clearance andincreased Mean Residence Time compared to the non-adapted molecule.Increased half-life can improve the pharmacokinetic and pharmacodynamicproperties of a therapeutic molecule and can also be important forimproved patient compliance.

The phrases, “half-life” (“t_(1/2)”) and “serum half life”, refer to thetime taken for the serum (or plasma) concentration of an antigen bindingprotein in accordance with the disclosure to reduce by 50%, in vivo, forexample due to degradation of the antigen binding protein and/orclearance or sequestration of the antigen binding protein by naturalmechanisms.

The antigen binding proteins of the disclosure can be stabilized in vivoand their half-life increased by binding to molecules which resistdegradation and/or clearance or sequestration (“half-life extendingmoiety” or “half-life extending molecule”). Half-life extensionstrategies are reviewed, for example, in “Therapeutic Proteins:Strategies to Modulate Their Plasma Half-Lives”, Edited by RolandKontermann, Wiley-Blackwell, 2012, ISBN: 978-3-527-32849-9. Suitablehalf-life extension strategies include: PEGylation, polysialylation,HESylation, recombinant PEG mimetics, N-glycosylation, O-glycosylation,Fc fusion, engineered Fc, IgG binding, albumin fusion, albumin binding,albumin coupling and nanoparticles.

In one embodiment, the half-life extending moiety or molecule is apolyethylene glycol moiety or a PEG mimetic. In one embodiment, theantigen binding protein comprises (optionally consists of) a singlevariable domain of the disclosure linked to a polyethylene glycol moiety(optionally, wherein said moiety has a size of about 20 to about 50 kDa,optionally about 40 kDa linear or branched PEG). Reference is made toWO04081026 for more detail on PEGylation of domain antibodies andbinding moieties. In one embodiment, the antagonist consists of a domainantibody monomer linked to a PEG, wherein the domain antibody monomer isa single variable domain according to the disclosure. Suitable PEGmimetics are reviewed, for example in Chapter 4, pages 63-80,“Therapeutic Proteins: Strategies to Modulate Their Plasma Half-Lives”Edited by Roland Kontermann, Wiley-Blackwell, 2012, ISBN:978-3-527-32849-9.

The interaction between the Fc region of an antibody and various Fcreceptors (FcγR) is believed to mediate phagocytosis andhalf-life/clearance of an antibody or antibody fragment. The neonatalFcRn receptor is believed to be involved in both antibody clearance andthe transcytosis across tissues (see Junghans (1997) Immunol. Res 16:29-57; and Ghetie et al. (2000) Annu. Rev. Immunol. 18: 739-766). In oneembodiment, the half-life extending moiety may be an Fc region from anantibody. Such an Fc region may incorporate various modificationsdepending on the desired property. For example, a salvage receptorbinding epitope may be incorporated into the antibody to increase serumhalf life, see U.S. Pat. No. 5,739,277.

Human IgG1 residues determined to interact directly with human FcRnincludes Ile253, Ser254, Lys288, Thr307, Gln311, Asn434 and His435.Accordingly, substitutions at any of the positions described in thissection may enable increased serum half-life and/or altered effectorproperties of the antibodies.

Half-life extension by fusion to the Fc region is reviewed, for example,in Chapter 9, pages 157-188, “Therapeutic Proteins: Strategies toModulate Their Plasma Half-Lives” Edited by Roland Kontermann,Wiley-Blackwell, 2012, ISBN: 978-3-527-32849-9.

Typically, a polypeptide that enhances serum half-life in vivo, i.e. ahalf-life extending molecule, is a polypeptide which occurs naturally invivo and which resists degradation or removal by endogenous mechanismswhich remove unwanted material from the organism (e.g., human).Typically, such molecules are naturally occurring proteins whichthemselves have a long half-life in vivo.

For example, a polypeptide that enhances serum half-life in vivo can beselected from proteins from the extracellular matrix, proteins found inblood, proteins found at the blood brain barrier or in neural tissue,proteins localized to the kidney, liver, muscle, lung, heart, skin orbone, stress proteins, disease-specific proteins, or proteins involvedin Fc transport. Suitable polypeptides are described, for example, inWO2008/096158.

Such an approach can also be used for targeted delivery of an antigenbinding protein, e.g. a single variable domain, in accordance with thedisclosure to a tissue of interest. In one embodiment targeted deliveryof a high affinity single variable domain in accordance with thedisclosure is provided.

In one embodiment, an antigen binding protein, e.g. single variabledomain, in accordance with the disclosure can be linked, i.e. conjugatedor associated, to serum albumin, fragments and analogues thereof.Half-life extension by fusion to albumin is reviewed, for example inChapter 12, pages 223-247, “Therapeutic Proteins: Strategies to ModulateTheir Plasma Half-Lives” Edited by Roland Kontermann, Wiley-Blackwell,2012, ISBN: 978-3-527-32849-9.

Examples of suitable albumin, albumin fragments or albumin variants aredescribed, for example, in WO2005077042 and WO2003076567.

In another embodiment, a single variable domain, polypeptide or ligandin accordance with the disclosure can be linked, i.e. conjugated orassociated, to transferrin, fragments and analogues thereof.

In one embodiment, half-life extension can be achieved by targeting anantigen or epitope that increases half-live in vivo. The hydrodynamicsize of an antigen binding protein and its serum half-life may beincreased by conjugating or associating an antigen binding protein ofthe disclosure to a binding domain that binds a naturally occurringmolecule and increases half-live in vivo.

For example, the antigen binding protein in accordance with theinvention can be conjugated or linked to an anti-serum albumin oranti-neonatal Fc receptor antibody or antibody fragment, e.g. an anti-SAor anti-neonatal Fc receptor dAb, Fab, Fab′ or scFv, or to an anti-SAaffibody or anti-neonatal Fc receptor Affibody or an anti-SA avimer, oran anti-SA binding domain which comprises a scaffold selected from, butnot limited to, the group consisting of CTLA-4, lipocallin, SpA, anaffibody, an avimer, GroEl and fibronectin (see WO2008096158 fordisclosure of these binding domains). Conjugating refers to acomposition comprising polypeptide, dAb or antagonist of the disclosurethat is bonded (covalently or noncovalently) to a binding domain such asa binding domain that binds serum albumin.

In another embodiment, the binding domain may be a polypeptide domainsuch as an Albumin Binding Domain (ABD) or a small molecule which bindsalbumin (reviewed, for example in Chapter 14, pages 269-283 and Chapter15, pages 285-296, “Therapeutic Proteins: Strategies to Modulate TheirPlasma Half-Lives” Edited by Roland Kontermann, Wiley-Blackwell, 2012,ISBN: 978-3-527-32849-9).

In one embodiment, there is provided a fusion protein comprising anantigen binding protein in accordance with the invention and ananti-serum albumin or anti-neonatal Fc receptor antibody or antibodyfragment.

The long half-life of IgG antibodies is reported to be dependent on itsbinding to FcRn. Therefore, substitutions that increase the bindingaffinity of IgG to FcRn at pH 6.0 while maintaining the pH dependence ofthe interaction by engineering the constant region have been extensivelystudied (Ghetie et al., Nature Biotech. 15: 637-640, 1997; Hinton etal., JBC 279: 6213-6216, 2004; Dall'Acqua et al.,

10 J Immunol 117: 1129-1138, 2006). Another means of modifying antigenbinding proteins of the present invention involves increasing thein-vivo half life of such proteins by modification of the immunoglobulinconstant domain or FcRn (Fc receptor neonate) binding domain.

In adult mammals, FcRn, also known as the neonatal Fc receptor, plays akey role in maintaining serum antibody levels by acting as a protectivereceptor that binds and salvages antibodies of the IgG isotype fromdegradation. IgG molecules are endocytosed by endothelial cells, and ifthey bind to FcRn, are recycled out into circulation. In contrast, IgGmolecules that do not bind to FcRn enter the cells and are targeted tothe lysosomal pathway where they are degraded.

The neonatal FcRn receptor is believed to be involved in both antibodyclearance and the transcytosis across tissues (see Junghans R. P (1997)Immunol. Res 16. 29-57 and Ghetie et al (2000) Annu. Rev. Immunol. 18,739-766). Human IgG1 residues determined to interact directly with humanFcRn includes Ile253, Ser254, Lys288, Thr307, Gln311, Asn434 and His435.Switches at any of these positions described in this section may enableincreased serum half-life and/or altered effector properties of antigenbinding proteins of the invention.

Antigen binding proteins of the present invention may have amino acidmodifications that increase the affinity of the constant domain orfragment thereof for FcRn. Increasing the half-life of therapeutic anddiagnostic IgG's and other bioactive molecules has many benefitsincluding reducing the amount and/or frequency of dosing of thesemolecules. In one embodiment there is therefore provided an antigenbinding according to the invention provided herein or a fusion proteincomprising all or a portion (an FcRn binding portion) of an IgG constantdomain having one or more of these amino acid modifications and anon-IgG protein or non-protein molecule conjugated to such a modifiedIgG constant domain, wherein the presence of the modified IgG constantdomain increases the in vivo half life of the antigen binding protein.

PCT Publication No. WO 00/42072 discloses a polypeptide comprising avariant Fc region with altered FcRn binding affinity, which polypeptidecomprises an amino acid modification at any one or more of amino acidpositions 238, 252, 253, 254, 255, 256, 265, 272, 286, 288, 303, 305,307, 309, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 386, 388,400, 413, 415, 424, 433, 434, 435, 436, 439, and 447 of the Fc region,wherein the numbering of the residues in the Fc region is that of the EUindex (Kabat et al).

PCT Publication No. WO 02/060919 A2 discloses a modified IgG comprisingan IgG constant domain comprising one or more amino acid modificationsrelative to a wild-type IgG constant domain, wherein the modified IgGhas an increased half-life compared to the half-life of an IgG havingthe wild-type IgG constant domain, and wherein the one or more aminoacid modifications are at one or more of positions 251, 253, 255,285-290, 308-314, 385-389, and 428-435.

Shields et al. (2001, J Biol Chem; 276:6591-604) used alanine scanningmutagenesis to alter residues in the Fc region of a human IgG1 antibodyand then assessed the binding to human FcRn. Positions that effectivelyabrogated binding to FcRn when changed to alanine include I253, S254,H435, and Y436. Other positions showed a less pronounced reduction inbinding as follows: E233-G236, R255, K288, L309, S415, and H433. Severalamino acid positions exhibited an improvement in FcRn binding whenchanged to alanine; notable among these are P238, T256, E272, V305,T307, Q311, D312, K317, D376, E380, E382, S424, and N434. Many otheramino acid positions exhibited a slight improvement (D265, N286, V303,K360, Q362, and A378) or no change (S239, K246, K248, D249, M252, E258,T260, S267, H268, S269, D270, K274, N276, Y278, D280, V282, E283, H285,T289, K290, R292, E293, E294, Q295, Y296, N297, S298, R301, N315, E318,K320, K322, S324, K326, A327, P329, P331, E333, K334, T335, S337, K338,K340, Q342, R344, E345, Q345, Q347, R356, M358, T359, K360, N361, Y373,S375, S383, N384, Q386, E388, N389, N390, K392, L398, S400, D401, K414,R416, Q418, Q419, N421, V422, E430, T437, K439, S440, S442, S444, andK447) in FcRn binding.

The most pronounced effect was found for combination variants withimproved binding to FcRn. At pH 6.0, the E380A/N434A variant showed over8-fold better binding to FcRn, relative to native IgG1, compared with2-fold for E380A and 3.5-fold for N434A. Adding T307A to this effected a12-fold improvement in binding relative to native IgG1. In oneembodiment the antigen binding protein of the invention comprises theE380A/N434A mutations and has increased binding to FcRn.

Dall'Acqua et al. (2002, J Immunol.; 169:5171-80) described randommutagenesis and screening of human IgG1 hinge-Fc fragment phage displaylibraries against mouse FcRn. They disclosed random mutagenesis ofpositions 251, 252, 254-256, 308, 309, 311, 312, 314, 385-387, 389, 428,433, 434, and 436. The major improvements in IgG1-human FcRn complexstability occur in substituting residues located in a band across theFc-FcRn interface (M252, S254, T256, H433, N434, and Y436) and to lesserextend substitutions of residues at the periphery like V308, L309, Q311,G385, Q386, P387, and N389. The variant with the highest affinity tohuman FcRn was obtained by combining the M252Y/S254T/T256E andH433K/N434F/Y436H mutations and exhibited a 57-fold increase in affinityrelative to the wild-type IgG1. The in vivo behaviour of such a mutatedhuman IgG1 exhibited a nearly 4-fold increase in serum half-life incynomolgus monkey as compared to wild-type IgG1.

The present invention therefore provides a variant of an antigen bindingprotein with optimized binding to FcRn. In a preferred embodiment, thesaid variant of an antigen binding protein comprises at least one aminoacid modification in the Fc region of said antigen binding protein,wherein said modification is selected from the group consisting of 226,227, 228, 230, 231, 233, 234, 239, 241, 243, 246, 250, 252, 256, 259,264, 265, 267, 269, 270, 276, 284, 285, 288, 289, 290, 291, 292, 294,297, 298, 299, 301, 302, 303, 305, 307, 308, 309, 311, 315, 317, 320,322, 325, 327, 330, 332, 334, 335, 338, 340, 342, 343, 345, 347, 350,352, 354, 355, 356, 359, 360, 361, 362, 369, 370, 371, 375, 378, 380,382, 384, 385, 386, 387, 389, 390, 392, 393, 394, 395, 396, 397, 398,399, 400, 401 403, 404, 408, 411, 412, 414, 415, 416, 418, 419, 420,421, 422, 424, 426, 428, 433, 434, 438, 439, 440, 443, 444, 445, 446 and447 of the Fc region as compared to said parent polypeptide, wherein thenumbering of the amino acids in the Fc region is that of the EU index inKabat.

In a further aspect of the invention the modifications areM252Y/S254T/T256E.

Additionally, various publications describe methods for obtainingphysiologically active molecules whose half-lives are modified either byintroducing an FcRn-binding polypeptide into the molecules (WO 97/43316;U.S. Pat. No. 5,869,046; U.S. Pat. No. 5,747,035; WO 96/32478; WO91/14438) or by fusing the molecules with antibodies whose FcRn-bindingaffinities are preserved but affinities for other Fc receptors have beengreatly reduced (WO 99/43713) or fusing with FcRn binding domains ofantibodies (WO 00/09560; U.S. Pat. No. 4,703,039).

Although substitutions in the constant region are able to significantlyimprove the functions of therapeutic IgG antibodies, substitutions inthe strictly conserved constant region have the risk of immunogenicityin human (Presta, supra, 2008; De Groot and Martin, Clin Immunol 131:189-201, 2009) and substitution in the highly diverse variable regionsequence might be less immunogenic. Reports concerned with the variableregion include engineering the CDR residues to improve binding affinityto the antigen (Rothe et al., Expert Opin Bioi Ther 6: 177-187, 2006;Bostrom et al., Methods Mol Bioi 525: 353-376, 2009; Thie et al.,Methods Mol Bioi 525: 309-322, 2009) and engineering the CDR andframework residues to improve stability (Worn and Pluckthun, J Mol Bioi305: 989-1010, 2001; Ewert et al., Methods 34: 184-199, 2004) anddecrease immunogenicity risk (De Groot and Martin, supra, 2009; Jones etal., Methods Mol Bio 525: 405-423, xiv, 2009). As reported, improvedaffinity to the antigen can be achieved by affinity maturation using thephage or ribosome display of a randomized library.

Improved stability can be rationally obtained from sequence- andstructure-based rational design. Decreased immunogenicity risk(deimmunization) can be accomplished by various humanizationmethodologies and the removal of T-cell epitopes, which can be predictedusing in silico technologies or determined by in vitro assays.Additionally, variable regions have been engineered to lower pI. Alonger half life was observed for these antibodies as compared to wildtype antibodies despite comparable FcRn binding. Engineering orselecting antibodies with pH dependent antigen binding to modifyantibody and/or antigen half life eg IgG2 antibody half life can beshortened if antigen-mediated clearance mechanisms normally degrade theantibody when bound to the antigen. Similarly, the antigen:antibodycomplex can impact the half-life of the antigen, either extendinghalf-life by protecting the antigen from the typical degradationprocesses, or shortening the half-life via antibody-mediateddegradation. One embodiment relates to antibodies with higher affinityfor antigen at pH 7.4 as compared to endosomal pH (i.e., pH 5.5-6.0)such that the KD ratio at pH5.5/pH 7.4 or at pH 6.0/pH 7.4 is 2 or more.For example to enhance the pharmacokinetic (PK) and pharmacodynamic (PD)properties of the antibody, it is possible to engineer pH-sensitivebinding to the antibody by introducing histidines into CDR residues.

Pharmaceutical Compositions

The antigen binding proteins of the present invention will normally, butnot necessarily, be formulated into pharmaceutical compositions prior toadministration to a patient. Accordingly, in another aspect of theinvention there is provided a pharmaceutical composition comprising anantigen binding protein and optionally one or more pharmaceuticallyacceptable excipients and/or carriers.

Methods for the preparation of such pharmaceutical compositions are wellknown to those skilled in the art (e.g. Remingtons PharmaceuticalSciences, 16th edition (1980) Mack Publishing Co and PharmaceuticalBiotechnology; Plenum publishing corporation; Volumes 2, 5 and 9).

The antigen binding proteins of the present invention may be formulatedfor administration in any convenient way. Pharmaceutical compositionsmay, for example, be administered by injection or continuous infusion(examples include, but are not limited to, intravenous, intraperitoneal,intradermal, subcutaneous, intramuscular and intraportal). In oneembodiment, the composition is suitable for subcutaneous injection.

Pharmaceutical compositions may be suitable for topical administration(which includes, but is not limited to, epicutaneous, inhaled,intranasal or ocular administration) or enteral administration (whichincludes, but is not limited to, oral or rectal administration).

Pharmaceutical compositions may comprise between 0.0001 mg/kg to 10mg/kg of antigen binding protein, for example between 0.1 mg/kg and 5mg/kg of antigen binding protein. Alternatively, the composition maycomprise between 1.0 mg/kg and 3.0 mg/kg.

Pharmaceutical compositions may comprise, in addition to an antigenbinding protein of the present invention, one or more other therapeuticagents.

Methods of Use

The antigen binding proteins described herein may have use in therapy.Antigen binding proteins of the present invention that bind LymphocyteActivation Gene 3 (LAG-3) and cause depletion of LAG-3+ activated Tcells may have use in the treatment or prevention of diseases associatedwith the involvement of pathogenic T cells, such as auto-immune diseasesand cancer.

Examples of disease states in which the antigen binding proteins havepotentially beneficial effects include psoriasis, Crohn's disease,rheumatoid arthritis, primary biliary cirrhosis, SLE, Sjögren'ssyndrome, multiple sclerosis, ulcerative colitis and autoimmunehepatitis.

The antigen binding proteins of the present invention may also have usein the treatment of cancer.

It will be appreciated by those skilled in the art that referencesherein to “treatment” or “therapy” may, depending on the condition,extend to prophylaxis in addition to the treatment of an establishedcondition.

There is thus provided as a further aspect of the invention an antigenbinding protein of the present invention for use in therapy.

There is also therefore provided an antigen binding protein of thepresent invention for use in the treatment of psoriasis, Crohn'sdisease, rheumatoid arthritis, primary biliary cirrhosis, SLE, Sjögren'ssyndrome, multiple sclerosis, ulcerative colitis or autoimmunehepatitis.

In a further embodiment, there is provided an antigen binding protein ofthe present invention for use in the treatment of psoriasis.

There is further provided the use of an antigen binding protein of thepresent invention in the manufacture of a medicament for the treatmentof psoriasis, Crohn's disease, rheumatoid arthritis, primary biliarycirrhosis, SLE, Sjögren's syndrome, multiple sclerosis, ulcerativecolitis or autoimmune hepatitis.

In a further embodiment, there is provided the use of an antigen bindingprotein of the present invention in the manufacture of a medicament forthe treatment of psoriasis.

There is further provided a method of treatment of psoriasis, Crohn'sdisease, rheumatoid arthritis, primary biliary cirrhosis, SLE, Sjögren'ssyndrome, multiple sclerosis, ulcerative colitis or autoimmunehepatitis, which method comprises administering to a human subject inneed thereof, a therapeutically effective amount of an antigen bindingprotein of the present invention.

In a further embodiment, there is provided a method of treatment ofpsoriasis, which method comprises administering to a human subject inneed thereof, a therapeutically effective amount of an antigen bindingprotein of the present invention.

The phrase “therapeutically effective amount” as used herein is anamount of an antigen binding protein of the present invention requiredto ameliorate or reduce one or more symptoms of, or to prevent or cure,the disease.

EXAMPLES

The following Examples illustrate but do not limit the invention.

Example 1 Biacore™ SPR Analysis of Purified Anti-LAG-3 HumanisedAntibodies

Anti-LAG-3 humanised antibodies were expressed in HEK 293 6E cells andpurified by affinity chromatography as follows:

100 ml Scale HEK 293 6E Expression

Expression plasmids encoding heavy and light chains of the humanisedantibodies identified in Table 3 were transiently co-transfected intoHEK 293 6E cells and expressed at 100 ml scale to produce antibody.

Purification of Humanised Antibodies

The expressed antibody molecules were purified by affinitychromatography. Protein A sepharose beads (GE Healthcare Cat No17-5280-04), were added to the cell culture supernatants and mixed atroom temperature for 30-60 minutes. The protein A sepharose beads werethen centrifuged and washed in PBS. The mixture was then added into a 10ml disposable column (Pierce Cat No: 29924) and the PBS allowed todrain. The column was washed 3 times with 10 ml PBS before elution ofthe antibody with IgG Elution buffer (Pierce Cat No: 21009). Theantibody eluted was immediately neutralized using 1M Trizma®hydrochloride buffer (T2819) and was then buffer exchanged into PBS witha Vivaspin 6 mwco 5000 column (Cat. No.: FDP-875-105D). The yield wasdetermined by measurement of absorbance at 280 nm. The level ofaggregated protein in the purified sample was determined by sizeexclusion chromatography.

Binding Kinetics

The binding kinetics of the purified antibodies for sLAG-3 were assessedby SPR using a Biacore™ 3000. A goat anti-human kappa antibody (SouthernBiotech, Catalogue No. 2060-01) was immobilised on a CM5 chip by primaryamine coupling. Humanised anti-LAG3 antibodies were captured on thissurface and the LAG3-Ig molecule IMP321 (Chrystelle Brignone, CarolineGrygar, Manon Marcu, Knut Schakel, Frederic Triebel, The Journal ofImmunology, 2007, 179: 4202-4211) used as the analyte at 64 nM with abuffer injection (i.e. 0 nM) used to double reference the bindingcurves. Regeneration was with 10 mM Glycine, pH1.5. The run was carriedout on the Biacore™ 3000, using HBS-EP as running buffer and at 25° C.Sensorgrams were normalised for binding at maximal association andoff-rates compared against IMP731 by visual inspection of the sensorgramprofiles.

The results show that the purified humanised anti-LAG-3 antibodies canbe categorised into 3 groups; group 1 with humanised variants thatappear to be better at binding to IMP321 than the chimeric IMP731, group2 with variants that appear to bind in a very similar manner to IMP731and group 3 with variants that demonstrate worse binding than IMP731.

10 humanised variants fall within group 1 and demonstrate improvedoff-rates when compared to IMP731 by visual inspection of the Biacore™sensorgrams. All of these molecules contained the L7 light chain, whichcontains a glycine to proline substitution at position 27e in lightchain CDR1. This data indicates that proline at position 27e improvesthe off-rate for IMP321 of the humanised anti-LAG-3 antibodies whenpaired with numerous humanised heavy chains.

Of the remaining antibodies tested, 4 molecules fall within group 2 andhad similar off-rates to the chimeric IMP731 antibody, while 4 othermolecules fall within group 3 and exhibit worse off-rates than IMP731.Table 3 provides an approximate ranking of those molecules in comparisonto IMP731. H5L7 exhibits the best off-rate in this experiment.

TABLE 3 Comparison of off-rates with IMP731 Off-rate Humanised Variantcomparison with IMP731 Rank order of off-rates H5L7 better 1 H1L7 better2 J7L7 better 3 H4L7 better 4 J11L7 better 5 H2L7 better 6 J13L7 better7 H7L7 better 8 J0L7 better 9 H0L7 better 10 H1L1 same H5L1 same J7L1same J11L1 same J13L1 worse H7L1 worse J0L1 worse H0L1 worse

Example 2 Binding Analysis of Wild Type Fucosvlated and AfucosylatedHumanised Anti-LAG-3 Antibodies for Binding to Recombinant HumanLAG-3-his Using the Biacore™ T100

Afucosylated antibody H5L7BW was generated by expression of plasmidsencoding H5L7 in the BioWa Potelligent® (FUT8 knock-out CHO) cell line.Afucosylated antibodies produced using Potelligent® technology have beenshown to exhibit increased antibody dependent cell-mediated cytotoxicity(ADCC) compared to equivalent highly-fucosylated conventional antibodiesthrough increased affinity to FcγRIIIa (CD16).

Wild type fucosylated and afucosylated antibodies were compared fortheir ability to bind to recombinant human LAG-3 ECD with a C-terminalHis6 (SEQ ID NO:51) using the Biacore™ T100 (GE Healthcare™). Protein Awas immobilised on a CM5 chip by primary amine coupling. This surfacewas then used to capture the humanised antibodies. Recombinant humanLAG-3 ECD-His6 was then passed over the captured antibodies at 32, 8, 2,0.5 and 0.125 nM and regeneration was carried out using 50 mM NaOH. Thebinding curves were double referenced with buffer injection (i.e. 0 nM)and the data was fitted to the T100 analysis software using the 1:1model. The run was carried out at 25° C., using HBS-EP as the runningbuffer.

In order to investigate the statistical significance of this kineticdata, this experiment was repeated 3 times with the same batch of thefour antibodies and the chimeric control. KD, ka and kd parameters wereeach log (base 10) transformed prior to separate statistical analyses.For each parameter, a mixed model analysis of variance (‘Anova’) wasperformed on transformed data, including terms for Run (random blockeffect) and Antibody.

From the Anova, geometric means were predicted for each antibody, alongwith statistically plausible ranges (95% confidence intervals) for eachmean. Planned comparisons of antibodies were performed within the Anova.Comparisons are presented as ratios of the two antibodies compared,again with 95% confidence intervals and p values. The ratios may also beinterpreted as a fold change, e.g. a ratio of 0.1 corresponds to a 90%decrease.

Geometric means were derived for each of the humanised antibodies andthe chimeric control IMP731 for the binding affinity (KD), shown inTable 4. The mean KD for H5L7 was 0.2075 nM, a significant decrease of92% (i.e. 10 fold decrease) with p<0.0001 when compared with IMP731.IMP731 exhibited a mean KD of 2.76 nM. Afucosylated H5L7BW exhibitsequivalent binding as fucosylated H5L7, with a geometric KD of 0.2179nM, with no significant difference (p=0.1790).

The improvement in affinity observed for H5L7 in comparison to IMP731 ispredominantly driven by differences in the off-rates of the antibodiesfor LAG-3-ECD-His6, shown in Table 5. There is an approximate 85%decrease (i.e. almost 10 fold decrease) in kd for H5L7 in comparisonIMP731 which is highly statistically significant. There is nosignificant difference between afucosylated H5L7BW and H5L7 (p=0.4408).

TABLE 4 Statistical evaluation for the KD of fucosylated andafucosylated anti-LAG-3 variants binding to recombinant human LAG-3-HisGeometric Means for KD (nM) Geometric Antibody Batch No mean Lower 95%CI Upper 95% CI H5L7 GRITS42382 2.08E−10 1.89E−10 2.28E−10 H5L7BWGRITS42760 2.18E−10 1.98E−10 2.40E−10 IMP731 020909 2.76E−09 2.51E−093.04E−09 Comparisons of KD KD comparison Ratio Lower 95% CI Upper 95% CIP value H5L7BW-H5L7 1.0502 0.97268 1.13389 0.179 H5L7-IMP731 0.077360.0715 0.0837 <.0001

TABLE 5 Statistical evaluation for the kd of fucosylated andafucosylated anti-LAG-3 variants binding to recombinant human LAG-3-HisGeometric Means for kd (1/s) Geometric Antibody Batch No mean Lower 95%CI Upper 95% CI H5L7 GRITS42382 3.95E−03 3.16E−03 4.94E−03 H5L7BWGRITS42760 4.41E−03 3.53E−03 5.51E−03 IMP731 020909 2.75E−02 2.20E−023.44E−02 Comparisons of kd KD comparison Ratio Lower 95% CI Upper 95% CIP value H5L7BW-H5L7 1.11715 0.81531 1.53073 0.4408 H5L7-IMP731 0.154780.11157 0.21471 <.0001

Example 3 Evaluation of the LAG-3-His Binding Epitope of Anti-LAG-3Humanised Variants in Comparison with Chimeric Antibody IMP731 Using theProteOn™

An epitope binning experiment was performed with H5L7 to evaluatewhether the LAG-3 epitope within which IMP731 binds was conserved uponhumanisation. Furthermore, the binding epitope of fucosylated andafucosylated humanised variants was also compared.

Epitope binding was evaluated using the ProteOn™ XPR36 (BioRad™)biosensor machine, by assessing whether anti-LAG-3 antibodies were ableto simultaneously bind to LAG-3-His in complex with antibody captured onthe ProteOn™ chip surface. IMP731 and non-competitive murine antibody17B4 were utilised as controls in this assay.

The antibodies to be tested were biotinylated using Ez-Link-sulfo-NHSbiotinylation kit. Each biotinylated antibody was captured on a separateflow cell on a neutravidin NLC chip using a vertical injection. Afterantibody capture, the surface was blocked with biocytin at 2.5 mg/ml.Using the co-inject facility inherent to the ProteOn run software, LAG-3ECD-His6 was injected over the coupled antibodies at 100 nM, followed bythe un-biotinylated antibodies at 100 nM with both injections beinghorizontal so that the LAG-3-His and the antibodies cross all 6neutravidin captured antibodies. The assay was run at 25° C. and inHBS-EP on the ProteOn XPR36Protein Interaction Array System.

Data analysis was carried out using report points taken after theLAG-3-His injection and report points taken after the un-biotinylatedantibody analyte injection. The overall response was calculated bysubtracting the response seen with the antibody binding from theresponse seen with LAG-3 ECD-His6 binding. A positive resonance unit(RU) value meant that antibody analyte injection had bound to LAG-3ECD-His6 complexed with the biotinylated antibody on the neutravidincapture surface, indicative of binding at non-competitive epitopes. Noresponse or a negative response meant that antibody analyte injectionhad not bound to LAG-3 ECD-His6 complexed with the biotinylated antibodyon the neutravidin capture surface, indicating antibodies bind atcompetitive epitopes.

Anti-LAG-3 humanised variants (fucosylated and afucosylated) are unableto bind to human LAG-3 ECD-His6 in complex with IMP731, indicative thatthe epitope for LAG-3 ECD-His6 is shared and thus conserved. Murineantibody 17B4 is able to bind to human LAG-3 ECD-His6 in complex withIMP731 substantiating that this antibody is non-competitive for LAG-3ECD-His6 binding with IMP731. Conversely, the humanised antibodies areable to bind to human LAG-3 ECD-His6 in complex with 17B4.

The results confirm that there has been no alteration in epitope betweenthe chimeric IMP731 and the humanised variant of IMP731 tested in thisexperiment. The experiment also shows that there is no differencebetween fucosylated and afucosylated antibodies for binding.

Example 4 Binding Analysis of Anti-LAG-3 Humanised Antibodies toRecombinant Cynomolgus Macaque and Baboon LAG-3 ECD-His6 Using theBiacore™T100

The binding cross reactivity of anti-LAG-3 humanised antibodies for bothcynomolgus macaque (cyno) and baboon recombinant LAG-3 ECD-His6 (SEQ IDNOs 52 and 53, respectively) was assessed using the Biacore™ T100 (GEHealthcare™).

Protein A was immobilised on a CM5 chip by primary amine coupling. Thissurface was then used to capture the humanised antibodies. In-housegenerated recombinant cynomolgus macaque and baboon LAG-3 ECD-His6 werethen passed over the captured antibodies and regeneration was carriedout using 50 mM NaOH. LAG-3 ECD-His6 was passed over at 16, 4, 1, 0.25and 0.0625 nM. The binding curves were double referenced with bufferinjection (i.e. 0 nM) and the data was fitted to the T100 analysissoftware using the 1:1 model. The run was carried out at 37° C. usingHBS-EP as the running buffer.

H5L7, H5L7BW and IMP731 bind with comparable affinity to both cynomolgusmacaque and baboon recombinant LAG-3 ECD-His6. Data was generated fromseparate experiments, however chimeric antibody IMP731 was utilised as acontrol between experiments.

This data indicates that both non-human primate recombinant LAG-3orthologues bind to the H5L7 derived antibodies with a 10 foldimprovement in affinity in comparison with human recombinant LAG-3ECD-His6 (SEQ ID NO: 51) (H5L7: human LAG-3 0.208 nM, cyno LAG-3 0.017nM and baboon LAG-3 0.022 nM; H5L7BW human LAG-3 0.218 nM, cyno LAG-30.021 nM and baboon LAG-3 0.024 nM). Whilst both non-human primaterecombinant LAG-3 orthologues bind to IMP731 derived antibodies with animprovement in affinity of approximately 100 fold in comparison withhuman recombinant LAG-3 ECD-His6 (SEQ ID NO: 51) (IMP731: human LAG-32.76 nM, cyno LAG-3 0.021 nM and baboon LAG-3 0.019 nM).

Example 5 Binding Profiles of H5L7BW, H5L7 and IMP731 to Human LAG-3Expressing EL4 Cells, and Human Primary Activated CD3+ T Cells

Human LAG-3 expressing EL4 cells were incubated with either IMP731,H5L7WT or H5L7BW Alexa647-conjugated antibodies at varyingconcentrations of up to 50 μg/ml for 30 minutes at room temperature.Cells were washed with FACS buffer (PBS+0.5% BSA) to remove unboundantibody.

CD3⁺ T cells were prepared from PBMCs by negative selection usingUntouched Human T cell Dynal beads. CD3⁺ T cells were activated byincubation with immobilised anti-CD3 and anti-CD28 and solublerecombinant human IL-12 for 3 days at 37° C. Activated cells wereincubated with Alexa 647-conjugated antibodies of varying concentrationsup to 3 μg/ml for 30 minutes at room temperature. Cells were washed withFACS buffer (PBS+0.5% BSA).

EL4 cells and CD3⁺ T cells were analysed by FACS using a Beckman CoulterFC 500 flow cytometer. FACS raw data were analysed using CXP Analysissoftware (Beckman Coulter). The data was initially plotted on aforward-scatter versus side-scatter plot and the population of cells ofinterest were gated. This gated population was then plotted as ahistogram displaying fluorescence intensity and the mean fluorescenceintensity (MFI) calculated. MFI values were then plotted in GraphPadPrism 5 software to generate dose response curves.

Binding profiles of H5L7BW, H5L7 and IMP731 to human LAG-3 expressingEL4 cells, and human primary activated CD3+ T cells are shown in FIG. 1Aand FIG. 1B. The three antibodies exhibited similar bindingcharacteristics to human LAG-3 expressing EL4 cells (FIG. 1A) andactivated human CD3+ T cells (FIG. 1B).

Example 6 Assessment of the Depleting Activity of H5L7BW and H5L7 byAntibody Dependent Cellular Cytotoxicity (ADCC) in Primary Human T Cells

The donors used in these experiments were screened for re-call responsesto the CD4 antigens present in Revaxis, (Diptheria, Tetanus andPoliomyelitis) and to either a CMVpp65 peptide pool or to a CD8 peptidepool, which contained peptide epitopes to CMV, EBV and Influenza. Theantigen used to perform initial studies was the CMVpp65 peptide pool.However, due to the limited number of donors that demonstrated a re-callresponse to this antigen, Revaxis and the CD8 peptide pool were theantigens used to generate the majority of the potency data for theanti-LAG3 antibodies.

Peripheral blood was collected from healthy human volunteers on day 0(25 mL) and day 5 (75 mL) of the experiment and was used to preparemononuclear cells by ficollplaque density gradient centrifugation. ThePBMCs prepared on day 0 were labeled using the CellTrace™ Violet CellProliferation Kit, in accordance with the manufacturer's instructions,after which they were washed, seeded in 24-well flat bottomed tissueculture plates at a density of 2×10⁶/mL in medium and stimulated withantigen (Revaxis and the CD8 peptide pool were used at a dilution of 1in 1000; the CMVpp65 peptide was used at a dilution of 1 in 100). ThePBMCs were incubated for 5 days at 37° C. in a 5% CO2 humidifiedincubator.

Autologous donor NK cells were purified from the PBMCs prepared on day 5of the experiment by negative selection using kits from eitherInvitrogen or Miltenyi Biotec in accordance with the specificinstructions of each manufacturer. The purified NK cells were countedand diluted to a density of 1.3×106/mL in medium.

The cells that had been stimulated with antigen for 5 days were washed,counted and diluted to a density of 2×106/viable cells/mL in medium.

Autologous donor NK cells (80 μL) and antigen activated cells (100 μL)were pipetted into round-bottomed 5 mL polystyrene FACS tubes with testantibody or medium (20 μL). The final concentrations of the anti LAG-3antibodies tested ranged from 1000 to 0.015 ng/mL. The samples wereincubated for 18 h at 37° C. in a 5% CO2 humidified incubator. After 18h the ADCC assay samples were analysed for the presence of antigenspecific LAG-3 positive T cells using flow cytometry. Briefly, thesamples were washed in FACS buffer, blocked with human IgG (3 μg/tube),and incubated with mouse anti-human CD4, CD8, CD337, CD25 and LAG-3fluorochrome conjugated antibodies for 30 minutes in the dark at roomtemperature. After a further wash step in PBS the cells were incubatedin the presence of a fixable green dead cell for 30 minutes in the darkat room temperature. The samples were finally washed with PBS, fixed andanalysed by flow cytometry using a 60 second acquisition time at a flowrate of 1 μL per second.

All sample data were acquired using the BD FACSCanto II Flow Cytometerwith FACS Diva software version 6.1.3 (BD BioSciences). The gatingstrategy used is shown in FIG. 2. The Live/Dead fixable green dead cellstain was used as a dead cell exclusion marker and appropriate isotypeand FMO-1 controls were used to define negative populations. Brieflydead cells were omitted from the analysis using a plot of forwardscatter (FSC-A) against fluorescence in FL1 (Green dead cell stain).Doublets were then excluded from the analysis by using a plot of FSC-Wagainst FSC-H. Viable single lymphocytes were subsequently identifiedand gated using a plot of forward scatter (FSC-A) against side scatter(SSC-A). CD4 antigen specific T cells were identified from this gatedpopulation of cells using plots of CD4 fluorescence against SSC-A andViolet Dye fluorescence against CD4 fluorescence respectively. Antigenspecific CD4 T cells were identified by a reduction of Violet Dyefluorescence. A further plot of CD25 fluorescence against LAG-3fluorescence was drawn to confirm the activation state of thispopulation of cells and that they expressed LAG-3. Similar plots weredrawn to identify antigen specific CD8 T cells.

The percentages of antigen specific CD4 and CD8 T cells present in eachsample (as a percentage of the viable lymphocyte population) wererecorded. Nonlinear variable slope curve-fits of these data were plottedand EC50 values were generated using GraphPad Prism software (v5.03).

To calculate the maximum level of depletion observed in an assay, at thehighest concentration of antibody tested, the following formula wasused: (1=(% Antigen specific T cells remaining after antibodytreatment)/(% Antigen specific T cells remaining in the absence ofantibody))*100.

Potency data for the depletion of LAG3 positive antigen specific CD8 andCD4 T cells by H5L7BW and H5L7 was generated using the in vitro assaysystem detailed above. H5L7BW induced depletion of LAG3 positive antigenspecific CD4 and CD8 T cells by ADCC in six of the seven donors studiedfor disappearance of the respective cell types. The potency of H5L7BW inantigen specific CD4 T cells, as quantified by EC50 values, ranged from14 pg/mL to 3.4 ng/mL with maximum levels of depletion ranging from 44to 78%. The potency of this antibody in antigen specific LAG-3 positiveCD8 T cells ranged from 122 pg/mL to 17.5 ng/mL, with maximum levels ofdepletion ranging from 39 to 87%. H5L7 mediated low levels of depletion,or was inactive, in the donors studied.

Example 7 Assessment of the Depleting Activity of H5L7BW and H5L7 in anIn-Vivo Human PBMC/Mouse SCID Xenograft Model

This assay describes the use of a human PBMC/mouse SCID xenograft modelto assess in vivo depletion efficacy of the monoclonal, fully humanised,afucosylated LAG-3 depleting antibody H5L7BW on activated human T cells.Healthy volunteer peripheral blood mononuclear cells (PBMCs) wereisolated and stimulated overnight (anti-CD3, IL-12) to induce LAG-3expression prior to injection into the peritoneum of immuno-compromisedSCID mice. LAG-3 depleting or control antibodies were eitherco-administered into the peritoneum or injected intravenously.

Depletion of LAG-3 positive cells was assessed by flow cytometry inperitoneal lavage samples 5 or 24 hours after cell injection.

Mice were injected with activated huPBMCs (4×106-2×107 cells in 0.4 mlof PBS) by the intraperitoneal route. Depending on the particular studyroute, LAG-3 antibodies or huIgG1 BioWa controls were eitherco-administered i.p. with the huPBMCs or mice were pre-treatedintravenously 18 h prior to huPBMC injection. 5 or 24 hours post-cellinjection, mice were euthanized, a peritoneal lavage was performed andthe cellular content of the lavage buffer analysed by flow cytometry.Briefly, peritoneal lavage involved 3×5 ml washes of the intactperitoneal cavity using cold PBS containing 3 mM EDTA.

All sample data were acquired using the BD FACSCanto II Flow Cytometerwith FACS Diva software version 6.1.3 (BD BioSciences). Approximately1×106 cells (where possible) were added per FACS tube, the cellsuspensions centrifuged at 1500 rpm for 10 minutes, and the pellet thenre-suspended in 3 ml PBS. The supernatants were then carefully decantedand the cell pellets re-suspended in 100 μl cold FACS wash buffer. 15 μlFcR Blocking Reagent was then added per tube and the cells incubated for10 minutes at room temperature. 5 μl of each staining antibody (10 μl of1:100 pre-diluted anti-LAG-3 blocking/detection antibody 17B4-PE) orisotype control were then added respectively and incubated for 20minutes at room temperature protected from light. The cells weresubsequently washed by addition of 4 ml FACS buffer per tube andcentrifugation at 1500 rpm for 5 minutes after which the supernatant wascarefully decanted. This washing step was repeated. Finally the cellswere re-suspended in 300 μl FACS buffer and analysed by flow cytometry.In addition to LAG-3 detection by use of the fluorescently labeled LAG-3blocking antibody 17B4-PE, the following T cell and activation markerswere used for T cell phenotyping: CD45, CD4, CD8, and CD25. Percentagesof CD4 and CD8 T cells were expressed as percentage of CD45 positivecells. LAG-3 positive T cell populations were expressed as percentagesof their parent populations CD4 and CD8.

FACS Diva software version 6.1.3 was used to generate batch analyses ofindividual cell counts/events and cell population percentages. Data wereanalysed with SAS version 9.2.2 software using a generalized linearmodel for binomial data. This analysis directly models the cell count asa proportion of the parent population. In this way, the size of theparent population is taken into account during the analysis. Means werecalculated for proportions of target cell type for each treatment, alongwith 95% confidence intervals. These were expressed as percentages.

Planned comparisons of treatments versus Control were made using oddsratios. This expresses the odds of having a target cell type in onetreatment as a ratio to the odds of having the target cell type oncontrol treatment. An odds ratio<1 would indicate a reduced odds ofhaving the cell type in the treatment of interest compared to thecontrol. An odds ratio<1 would indicate a reduced odds of having thecell type in the treatment of interest compared to the control.

All experiments were performed using PBMCs from individual healthy blooddonors and due to the large blood volumes required per experiment, nodonor was used more than once. LAG-3 expression levels after overnightstimulation varied greatly between donors (ranging from 2-73% cellsurface expression) but these differences in expression levels had noeffect on the highly significant depletion efficacy of the testedantibodies.

The human PBMC/mouse SCID in vivo model was successfully used to showdepletion of activated human, LAG-3 expressing T cells in the peritoneumof immunocompromised SCID mice. Depending on donor, route ofadministration and time point of analysis, between 84.22-99.71% of LAG-3positive human CD4 T cells and between 84.64-99.62% of LAG-3 positivehuman CD8 T cells were depleted. As shown in FIG. 3A and FIG. 3B,co-administration of 5 mg/kg H5L7BW to activated human PBMCs in theperitoneum of SCID mice led to highly significant depletion of LAG-3positive CD4 and CD8 positive T cells 24 hours after injection comparedto Control IgG injected animals.

FIG. 4A and FIG. 4B highlight the comparison between 5 mg/kg H5L7BW andH5L7 5 hours after co-i.p. administration to activated human PBMCs asdescribed before. Both antibodies induced highly significant depletionof LAG-3 positive CD4 and CD8 T cells (FIG. 4A).

As shown in FIG. 5A and FIG. 5B, administration of 5 mg/kg H5L7BW viathe intra-venous route resulted in a highly statistically significantdepletion of LAG-3 positive T cells after 5 hours, similar to what wasobserved in the experiments with i.p. co-administered LAG-3 depletionantibodies (FIG. 5A). The comparison between i.v. administered H5L7BW,H5L7 and IMP731 (all at 5 mg/kg) 5 hours after i.p. administration ofactivated human PBMCs revealed very similar depletion efficacies betweenthe 3 molecules compared to control treated animals (FIG. 6A and FIG.6B). Each of the 3 tested LAG-3 depleting antibodies caused highlysignificant reduction in the number of LAG-3 positive CD4 and CD8 Tcells (FIG. 6A) with H5L7BW demonstrating greater depletion capacitycompared to H5L7 or IMP731.

Sequences

TABLE 6 Sequence Summary Sequence Identifier (SEQ ID No.) Amino acidPolynucleotide Sequence Sequence Sequence H5L7, CDRL1 (Kabat defined)  1 57 H5L7, CDRL2 (Kabat defined)  2  58 H5L7, CDRL3 (Kabat defined)  3 59 H5L7 VL  4  60 H5L7, light chain humanised  5  61 constructH5L7, CDRH1 (Kabat defined)  6  62 H5L7, CDRH2 (Kabat defined)  7  63H5L7, CDRH3 (Kabat defined)  8  64 H5L7 VH  9  65H5L7, heavy chain humanised 10  66 construct H1L7, light chain humanised11  67 construct H1L7, heavy chain humanised 12  68 constructJ7L7, light chain humanised 13  69 construct J7L7, heavy chain humanised14  70 construct H4L7, light chain humanised 15  71 constructH4L7, heavy chain humanised 16  72 constructJ11L7, light chain humanised 17  73 constructJ11L7, heavy chain humanised 18  74 constructH2L7, light chain humanised 19  75 construct H2L7, heavy chain humanised20  76 construct J13L7, light chain humanised 21  77 constructJ13L7, heavy chain humanised 22  78 constructH7L7, light chain humanised 23  79 construct H7L7, heavy chain humanised24  80 construct J0L7, light chain humanised 25  81 constructJ0L7, heavy chain humanised 26  82 construct H0L7, light chain humanised27  83 construct H0L7, heavy chain humanised 28  84 constructH1L1, light chain humanised 29  85 construct H1L1, heavy chain humanised30  86 construct H5L1, light chain humanised 31  87 constructH5L1, heavy chain humanised 32  88 construct J7L1, light chain humanised33  89 construct J7L1, heavy chain humanised 34  90 constructJ11L1, light chain humanised 35  91 constructJ11L1, heavy chain humanised 36  92 constructJ13L1, light chain humanised 37  93 constructJ13L1, heavy chain humanised 38  94 constructH7L1, light chain humanised 39  95 construct H7L1, heavy chain humanised40  96 construct J0L1, light chain humanised 41  97 constructJ0L1, heavy chain humanised 42  98 construct H0L1, light chain humanised43  99 construct H0L1, heavy chain humanised 44 100 constructHuman kappa chain constant 45 101 region Human IgG1 constant region 46102 IMP731, VH 47 103 IMP731, VL 48 104 IMP731, heavy chain sequence 49105 IMP731, light chain sequence 50 106 Recombinant human LAG-3- 51 107ECD-His6 Recombinant cynomolgus 52 108 macaque LAG-3 ECD-His6Recombinant baboon LAG-3 53 109 ECD-His6 Leader sequence used for 54 110humanised variant heavy and light chain constructsIMP731 Leader sequence 55 111 Leader sequence used for 56 112soluble LAG-3 constructs SEQ ID NO. 1 KSSQSLLNPSNQKNYLA SEQ ID NO. 2FASTRDS SEQ ID NO. 3 LQHFGTPPT SEQ ID NO. 4DIQMTQSPSSLSASVGDRVTITCKSSQSLLNPSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKR SEQ ID NO. 5DIQMTQSPSSLSASVGDRVTITCKSSQSLLNPSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 6 AYGVN SEQ ID NO. 7 MIWDDGSTDYDSALKS SEQ ID NO. 8 EGDVAFDYSEQ ID NO. 9QVQLQESGPGLVKPSETLSLTCTVSGFSLTAYGVNWIRQPPGKGLEWIGMIWDDGSTDYDSALKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREGDVAFDYWGQGTLVTVSS SEQ ID NO. 10QVQLQESGPGLVKPSETLSLTCTVSGFSLTAYGVNWIRQPPGKGLEWIGMIWDDGSTDYDSALKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 11DIQMTQSPSSLSASVGDRVTITCKSSQSLLNPSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 12QVQLQESGPGLVKPSETLSLTCTVSGFSLTAYGVNWIRQPPGKGLEWIGMIWDDGSTDYNSALKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 13DIQMTQSPSSLSASVGDRVTITCKSSQSLLNPSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 14QVQLVQSGAEVKKPGSSVKVSCKASGFSLTAYGVNWVRQAPGQGLEWMGMIWDDGSTDYNSALKSRVTITADKSTSTAYMELSSLRSEDTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 15DIQMTQSPSSLSASVGDRVTITCKSSQSLLNPSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 16QVQLQESGPGLVKPSETLSLTCTVSGFSLTAYGVNWIRQPPGKGLEWLGMIWDDGSTDYNSALKSRLTISKDNSKNQVSLKLSSVTAADTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 17DIQMTQSPSSLSASVGDRVTITCKSSQSLLNPSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 18QVQLVQSGAEVKKPGSSVKVSCKASGFSLTAYGVNWVRQAPGQGLEWMGMIWDDGSTDYDSALKSRVTITADKSTSTAYMELSSLRSEDTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 19DIQMTQSPSSLSASVGDRVTITCKSSQSLLNPSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 20QVQLQESGPGLVKPSETLSLTCTVSGFSLTAYGVNWIRQPPGKGLEWIGMIWDDGSTDYNSALKSRVTISKDNSKNQVSLKLSSVTAADTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 21DIQMTQSPSSLSASVGDRVTITCKSSQSLLNPSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 22QVQLVQSGAEVKKPGSSVKVSCKASGGTFSAYGVNWVRQAPGQGLEWMGMIWDDGSTDYDSALKSRVTITADKSTSTAYMELSSLRSEDTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 23DIQMTQSPSSLSASVGDRVTITCKSSQSLLNPSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 24QVQLQESGPGLVKPSETLSLTCTVSGGSISAYGVNWIRQPPGKGLEWIGMIWDDGSTDYDSALKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 25DIQMTQSPSSLSASVGDRVTITCKSSQSLLNPSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 26QVQLVQSGAEVKKPGSSVKVSCKASGGTFSAYGVNWVRQAPGQGLEWMGMIWDDGSTDYNSALKSRVTITADKSTSTAYMELSSLRSEDTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 27DIQMTQSPSSLSASVGDRVTITCKSSQSLLNPSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 28QVQLQESGPGLVKPSETLSLTCTVSGGSISAYGVNWIRQPPGKGLEWIGMIWDDGSTDYNSALKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 29DIQMTQSPSSLSASVGDRVTITCKSSQSLLNGSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 30QVQLQESGPGLVKPSETLSLTCTVSGFSLTAYGVNWIRQPPGKGLEWIGMIWDDGSTDYNSALKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 31DIQMTQSPSSLSASVGDRVTITCKSSQSLLNGSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 32QVQLQESGPGLVKPSETLSLTCTVSGFSLTAYGVNWIRQPPGKGLEWIGMIWDDGSTDYDSALKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 33DIQMTQSPSSLSASVGDRVTITCKSSQSLLNGSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 34QVQLVQSGAEVKKPGSSVKVSCKASGFSLTAYGVNWVRQAPGQGLEWMGMIWDDGSTDYNSALKSRVTITADKSTSTAYMELSSLRSEDTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 35DIQMTQSPSSLSASVGDRVTITCKSSQSLLNGSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 36QVQLVQSGAEVKKPGSSVKVSCKASGFSLTAYGVNWVRQAPGQGLEWMGMIWDDGSTDYDSALKSRVTITADKSTSTAYMELSSLRSEDTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 37DIQMTQSPSSLSASVGDRVTITCKSSQSLLNGSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 38QVQLVQSGAEVKKPGSSVKVSCKASGGTFSAYGVNWVRQAPGQGLEWMGMIWDDGSTDYDSALKSRVTITADKSTSTAYMELSSLRSEDTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 39DIQMTQSPSSLSASVGDRVTITCKSSQSLLNGSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 40QVQLQESGPGLVKPSETLSLTCTVSGGSISAYGVNWIRQPPGKGLEWIGMIWDDGSTDYDSALKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 41DIQMTQSPSSLSASVGDRVTITCKSSQSLLNGSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 42QVQLVQSGAEVKKPGSSVKVSCKASGGTFSAYGVNWVRQAPGQGLEWMGMIWDDGSTDYNSALKSRVTITADKSTSTAYMELSSLRSEDTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 43DIQMTQSPSSLSASVGDRVTITCKSSQSLLNGSNQKNYLAWYQQKPGKAPKLLVYFASTRDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQHFGTPPTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 44QVQLQESGPGLVKPSETLSLTCTVSGGSISAYGVNWIRQPPGKGLEWIGMIWDDGSTDYNSALKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREGDVAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 45TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO. 46ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 47QVQLKESGPGLVAPSQSLSITCTVSGFSLTAYGVNWVRQPPGKGLEWLGMIWDDGSTDYNSALKSRLSISKDNSKSQVFLKMNSLQTDDTARYYCAREGDVAFDYWGQGTTLTVSS SEQ ID NO. 48DIVMTQSPSSLAVSVGQKVTMSCKSSQSLLNGSNQKNYLAWYQQKPGQSPKLLVYFASTRDSGVPDRFIGSGSGTDFTLTISSVQAEDLADYFCLQHFGTPPTFGGGTKLEIKR SEQ ID NO. 49(Note different leader sequence used for chimeric antibodies)QVQLKESGPGLVAPSQSLSITCTVSGFSLTAYGVNWVRQPPGKGLEWLGMIWDDGSTDYNSALKSRLSISKDNSKSQVFLKMNSLQTDDTARYYCAREGDVAFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO. 50DIVMTQSPSSLAVSVGQKVTMSCKSSQSLLNGSNQKNYLAWYQQKPGQSPKLLVYFASTRDSGVPDRFIGSGSGTDFTLTISSVQAEDLADYFCLQHFGTPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO. 51LQPGAEVPVVWAQEGAPAQLPCSPTIPLQDLSLLRRAGVTWQHQPDSGPPAAAPGHPLAPGPHPAAPSSWGPRPRRYTVLSVGPGGLRSGRLPLQPRVQLDERGRQRGDFSLWLRPARRADAGEYRAAVHLRDRALSCRLRLRLGQASMTASPPGSLRASDWVILNCSFSRPDRPASVHWFRNRGQGRVPVRESPHHHLAESFLFLPQVSPMDSGPWGCILTYRDGFNVSIMYNLTVLGLEPPTPLTVYAGAGSRVGLPCRLPAGVGTRSFLTAKWTPPGGGPDLLVTGDNGDFTLRLEDVSQAQAGTYTCHIHLQEQQLNATVTLAIITVTPKSFGSPGSLGKLLCEVTPVSGQERFVWSSLDTPSQRSFSGPWLEAQEAQLLSQPWQCQLYQGERLLGAAVYFTELSSPHHHHHH SEQ ID NO. 52PQPGAEISVVWAQEGAPAQLPCSPTIPLQDLSLLRRAGVTWQHQPDSGPPAPAPGHPPAPGHRPAAPYSWGPRPRRYTVLSVGPGGLRSGRLPLQPRVQLDERGRQRGDFSLWLRPARRADAGEYRATVHLRDRALSCRLRLRVGQASMTASPPGSLRTSDWVILNCSFSRPDRPASVHWFRSRGQGRVPVQGSPHHHLAESFLFLPHVGPMDSGLWGCILTYRDGFNVSIMYNLTVLGLEPATPLTVYAGAGSRVELPCRLPPAVGTQSFLTAKWAPPGGGPDLLVAGDNGDFTLRLEDVSQAQAGTYICHIRLQGQQLNATVTLAIITVTPKSFGSPGSLGKLLCEVTPASGQEHFVWSPLNTPSQRSFSGPWLEAQEAQLLSQPWQCQLHQGERLLGAAVYFTELSSPHHHHHH SEQ ID NO. 53PQPGAEISVVWAQEGAPAQLPCSPTIPLQDLSLLRRAGVTWQHQPDSGPPAPAPGHPPAPGHRPAAPYSWGPRPRRYTVLSVGPGGLRSGRLPLQPRVQLDERGRQRGDFSLWLRPARRADAGEYRATVHLRDRALSCRLRLRVGQASMTASPPGSLRTSDWVILNCSFSRPDRPASVHWFRSRGQGQVPVQESPHHHLAESFLFLPHVGPMDSGLWGCILTYRDGFNVSIMYNLTVLGLEPTTPLTVYAGAGSRVELPCRLPPAVGTQSFLTAKWAPPGGGPDLLVVGDNGNFTLRLEDVSQAQAGTYICHIRLQGQQLNATVTLAIITVTPKSFGSPGSLGKLLCEVTPASGQERFVWSPLNTPSQRSFSGPWLEAQEAQLLSQPWQCQLHQGERLLGAAVYFTELSSPHHHHHH SEQ ID NO. 54MGWSCIILFLVATATGVHS SEQ ID NO. 55 MESQTQVLMFLLLWVSGACA SEQ ID NO. 56MPLLLLLPLL WAGALA SEQ ID NO. 57AAGAGCAGCCAGAGCCTGCTGAACCCCAGCAACCAGAAGAACTACCTGGCC SEQ ID NO. 58TTCGCCTCTACCAGGGATTCC SEQ ID NO. 59 CTGCAGCACTTCGGCACCCCTCCCACTSEQ ID NO. 60GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACCCCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGT SEQ ID NO. 61GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACCCCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 62 GCCTACGGCGTCAAC SEQ ID NO. 63ATGATCTGGGACGACGGCAGCACCGACTACGACAGCGCCCTGAAGAGC SEQ ID NO. 64GAGGGCGACGTGGCCTTCGATTAC SEQ ID NO. 65CAGGTGCAGCTCCAGGAGAGCGGCCCCGGCCTGGTGAAGCCTAGCGAGACCCTGAGCCTGACCTGCACCGTGAGCGGCTTCTCCCTGACCGCCTACGGCGTCAACTGGATCAGGCAGCCCCCCGGCAAAGGCCTGGAGTGGATTGGGATGATCTGGGACGACGGCAGCACCGACTACGACAGCGCCCTGAAGAGCAGGGTGACCATCAGCGTGGACACCAGCAAGAACCAGTTCAGCCTGAAGCTGAGCAGCGTGACTGCCGCCGACACCGCCGTCTATTACTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC SEQ ID NO. 66CAGGTGCAGCTCCAGGAGAGCGGCCCCGGCCTGGTGAAGCCTAGCGAGACCCTGAGCCTGACCTGCACCGTGAGCGGCTTCTCCCTGACCGCCTACGGCGTCAACTGGATCAGGCAGCCCCCCGGCAAAGGCCTGGAGTGGATTGGGATGATCTGGGACGACGGCAGCACCGACTACGACAGCGCCCTGAAGAGCAGGGTGACCATCAGCGTGGACACCAGCAAGAACCAGTTCAGCCTGAAGCTGAGCAGCGTGACTGCCGCCGACACCGCCGTCTATTACTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 67GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACCCCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 68CAGGTGCAGCTCCAGGAGAGCGGCCCCGGCCTGGTGAAGCCTAGCGAGACCCTGAGCCTGACCTGCACCGTGAGCGGCTTCTCCCTGACCGCCTACGGCGTCAACTGGATCAGGCAGCCCCCCGGCAAAGGCCTGGAGTGGATTGGGATGATCTGGGACGACGGCAGCACCGACTACAACAGCGCCCTGAAGAGCAGGGTGACCATCAGCGTGGACACCAGCAAGAACCAGTTCAGCCTGAAGCTGAGCAGCGTGACTGCCGCCGACACCGCCGTCTATTACTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 69GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACCCCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 70CAGGTGCAGCTCGTGCAGAGCGGGGCCGAAGTCAAGAAACCCGGCAGCTCCGTGAAGGTGAGCTGCAAGGCCAGCGGCTTCTCTCTCACTGCCTACGGCGTGAACTGGGTGAGGCAGGCTCCCGGCCAGGGCCTGGAGTGGATGGGCATGATCTGGGACGACGGCAGCACCGACTACAACAGCGCCCTGAAGAGCAGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGAGCGAGGACACCGCCGTGTACTATTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 71GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACCCCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 72CAGGTGCAGCTCCAGGAGAGCGGCCCCGGCCTGGTGAAGCCTAGCGAGACCCTGAGCCTGACCTGCACCGTGAGCGGCTTCTCCCTGACCGCCTACGGCGTCAACTGGATCAGGCAGCCCCCCGGCAAAGGCCTGGAGTGGCTGGGGATGATCTGGGACGACGGCAGCACCGACTACAACAGCGCCCTGAAGAGCAGGCTGACCATCAGCAAGGACAACAGCAAGAACCAGGTGAGCCTGAAGCTGAGCAGCGTGACTGCCGCCGACACCGCCGTCTATTACTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 73GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACCCCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 74CAGGTGCAGCTCGTGCAGAGCGGGGCCGAAGTCAAGAAACCCGGCAGCTCCGTGAAGGTGAGCTGCAAGGCCAGCGGCTTCTCTCTCACTGCCTACGGCGTGAACTGGGTGAGGCAGGCTCCCGGCCAGGGCCTGGAGTGGATGGGCATGATCTGGGACGACGGCAGCACCGACTACGACAGCGCCCTGAAGAGCAGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGAGCGAGGACACCGCCGTGTACTATTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 75GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACCCCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 76CAGGTGCAGCTCCAGGAGAGCGGCCCCGGCCTGGTGAAGCCTAGCGAGACCCTGAGCCTGACCTGCACCGTGAGCGGCTTCTCCCTGACCGCCTACGGCGTCAACTGGATCAGGCAGCCCCCCGGCAAAGGCCTGGAGTGGATTGGGATGATCTGGGACGACGGCAGCACCGACTACAACAGCGCCCTGAAGAGCAGGGTGACCATCAGCAAGGACAACAGCAAGAACCAGGTGAGCCTGAAGCTGAGCAGCGTGACTGCCGCCGACACCGCCGTCTATTACTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 77GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACCCCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 78CAGGTGCAGCTCGTGCAGAGCGGGGCCGAAGTCAAGAAACCCGGCAGCTCCGTGAAGGTGAGCTGCAAGGCCAGCGGCGGCACCTTCAGCGCCTACGGCGTGAACTGGGTGAGGCAGGCTCCCGGCCAGGGCCTGGAGTGGATGGGCATGATCTGGGACGACGGCAGCACCGACTACGACAGCGCCCTGAAGAGCAGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGAGCGAGGACACCGCCGTGTACTATTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 79GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACCCCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 80CAGGTGCAGCTCCAGGAGAGCGGCCCCGGCCTGGTGAAGCCTAGCGAGACCCTGAGCCTGACCTGCACCGTGAGCGGCGGCTCCATCAGCGCCTACGGCGTCAACTGGATCAGGCAGCCCCCCGGCAAAGGCCTGGAGTGGATTGGGATGATCTGGGACGACGGCAGCACCGACTACGACAGCGCCCTGAAGAGCAGGGTGACCATCAGCGTGGACACCAGCAAGAACCAGTTCAGCCTGAAGCTGAGCAGCGTGACTGCCGCCGACACCGCCGTCTATTACTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 81GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACCCCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 82CAGGTGCAGCTCGTGCAGAGCGGGGCCGAAGTCAAGAAACCCGGCAGCTCCGTGAAGGTGAGCTGCAAGGCCAGCGGCGGCACCTTCAGCGCCTACGGCGTGAACTGGGTGAGGCAGGCTCCCGGCCAGGGCCTGGAGTGGATGGGCATGATCTGGGACGACGGCAGCACCGACTACAACAGCGCCCTGAAGAGCAGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGAGCGAGGACACCGCCGTGTACTATTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 83GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACCCCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 84CAGGTGCAGCTCCAGGAGAGCGGCCCCGGCCTGGTGAAGCCTAGCGAGACCCTGAGCCTGACCTGCACCGTGAGCGGCGGCTCCATCAGCGCCTACGGCGTCAACTGGATCAGGCAGCCCCCCGGCAAAGGCCTGGAGTGGATTGGGATGATCTGGGACGACGGCAGCACCGACTACAACAGCGCCCTGAAGAGCAGGGTGACCATCAGCGTGGACACCAGCAAGAACCAGTTCAGCCTGAAGCTGAGCAGCGTGACTGCCGCCGACACCGCCGTCTATTACTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 85GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACGGCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 86CAGGTGCAGCTCCAGGAGAGCGGCCCCGGCCTGGTGAAGCCTAGCGAGACCCTGAGCCTGACCTGCACCGTGAGCGGCTTCTCCCTGACCGCCTACGGCGTCAACTGGATCAGGCAGCCCCCCGGCAAAGGCCTGGAGTGGATTGGGATGATCTGGGACGACGGCAGCACCGACTACAACAGCGCCCTGAAGAGCAGGGTGACCATCAGCGTGGACACCAGCAAGAACCAGTTCAGCCTGAAGCTGAGCAGCGTGACTGCCGCCGACACCGCCGTCTATTACTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 87GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACGGCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 88CAGGTGCAGCTCCAGGAGAGCGGCCCCGGCCTGGTGAAGCCTAGCGAGACCCTGAGCCTGACCTGCACCGTGAGCGGCTTCTCCCTGACCGCCTACGGCGTCAACTGGATCAGGCAGCCCCCCGGCAAAGGCCTGGAGTGGATTGGGATGATCTGGGACGACGGCAGCACCGACTACGACAGCGCCCTGAAGAGCAGGGTGACCATCAGCGTGGACACCAGCAAGAACCAGTTCAGCCTGAAGCTGAGCAGCGTGACTGCCGCCGACACCGCCGTCTATTACTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 89GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACGGCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 90CAGGTGCAGCTCGTGCAGAGCGGGGCCGAAGTCAAGAAACCCGGCAGCTCCGTGAAGGTGAGCTGCAAGGCCAGCGGCTTCTCTCTCACTGCCTACGGCGTGAACTGGGTGAGGCAGGCTCCCGGCCAGGGCCTGGAGTGGATGGGCATGATCTGGGACGACGGCAGCACCGACTACAACAGCGCCCTGAAGAGCAGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGAGCGAGGACACCGCCGTGTACTATTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 91GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACGGCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 92CAGGTGCAGCTCGTGCAGAGCGGGGCCGAAGTCAAGAAACCCGGCAGCTCCGTGAAGGTGAGCTGCAAGGCCAGCGGCTTCTCTCTCACTGCCTACGGCGTGAACTGGGTGAGGCAGGCTCCCGGCCAGGGCCTGGAGTGGATGGGCATGATCTGGGACGACGGCAGCACCGACTACGACAGCGCCCTGAAGAGCAGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGAGCGAGGACACCGCCGTGTACTATTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 93GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACGGCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 94CAGGTGCAGCTCGTGCAGAGCGGGGCCGAAGTCAAGAAACCCGGCAGCTCCGTGAAGGTGAGCTGCAAGGCCAGCGGCGGCACCTTCAGCGCCTACGGCGTGAACTGGGTGAGGCAGGCTCCCGGCCAGGGCCTGGAGTGGATGGGCATGATCTGGGACGACGGCAGCACCGACTACGACAGCGCCCTGAAGAGCAGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGAGCGAGGACACCGCCGTGTACTATTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 95GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACGGCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 96CAGGTGCAGCTCCAGGAGAGCGGCCCCGGCCTGGTGAAGCCTAGCGAGACCCTGAGCCTGACCTGCACCGTGAGCGGCGGCTCCATCAGCGCCTACGGCGTCAACTGGATCAGGCAGCCCCCCGGCAAAGGCCTGGAGTGGATTGGGATGATCTGGGACGACGGCAGCACCGACTACGACAGCGCCCTGAAGAGCAGGGTGACCATCAGCGTGGACACCAGCAAGAACCAGTTCAGCCTGAAGCTGAGCAGCGTGACTGCCGCCGACACCGCCGTCTATTACTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 97GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACGGCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 98CAGGTGCAGCTCGTGCAGAGCGGGGCCGAAGTCAAGAAACCCGGCAGCTCCGTGAAGGTGAGCTGCAAGGCCAGCGGCGGCACCTTCAGCGCCTACGGCGTGAACTGGGTGAGGCAGGCTCCCGGCCAGGGCCTGGAGTGGATGGGCATGATCTGGGACGACGGCAGCACCGACTACAACAGCGCCCTGAAGAGCAGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGAGCGAGGACACCGCCGTGTACTATTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 99GACATCCAGATGACCCAGAGCCCCTCTAGCCTCAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAAGAGCAGCCAGAGCCTGCTGAACGGCAGCAACCAGAAGAACTACCTGGCCTGGTACCAGCAGAAACCCGGCAAGGCCCCCAAGCTGCTGGTCTACTTCGCCTCTACCAGGGATTCCGGCGTCCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACACTGACCATCAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCTGCAGCACTTCGGCACCCCTCCCACTTTTGGCCAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCSEQ ID NO. 100CAGGTGCAGCTCCAGGAGAGCGGCCCCGGCCTGGTGAAGCCTAGCGAGACCCTGAGCCTGACCTGCACCGTGAGCGGCGGCTCCATCAGCGCCTACGGCGTCAACTGGATCAGGCAGCCCCCCGGCAAAGGCCTGGAGTGGATTGGGATGATCTGGGACGACGGCAGCACCGACTACAACAGCGCCCTGAAGAGCAGGGTGACCATCAGCGTGGACACCAGCAAGAACCAGTTCAGCCTGAAGCTGAGCAGCGTGACTGCCGCCGACACCGCCGTCTATTACTGCGCCAGGGAGGGCGACGTGGCCTTCGATTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 101ACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGC SEQ ID NO. 102GCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO. 103CAGGTGCAGCTGAAGGAGTCAGGTCCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCATCACATGCACCGTCTCAGGGTTCTCATTAACCGCCTATGGTGTAAACTGGGTTCGCCAGCCTCCAGGAAAGGGTCTGGAGTGGCTGGGAATGATATGGGATGATGGAAGCACAGACTATAATTCAGCTCTCAAATCCAGACTGAGCATCAGTAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCAGGTACTACTGTGCCAGAGAAGGGGACGTAGCCTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA SEQ ID NO. 104GACATTGTGATGACACAGTCTCCCTCCTCCCTGGCTGTGTCAGTAGGACAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGCCTTTTAAATGGTAGCAATCAAAAGAACTACTTGGCCTGGTACCAGCAGAAACCAGGACAGTCTCCTAAACTTCTGGTATACTTTGCATCCACTAGGGATTCTGGGGTCCCTGATCGCTTCATAGGCAGTGGATCTGGGACAGATTTCACTCTTACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGATTACTTCTGTCTGCAACATTTTGGCACTCCTCCGACGTTCGGTGGAGGCACCAAACTGGAAATCAAACGG SEQ ID NO. 105CAGGTGCAGCTGAAGGAGTCAGGTCCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCATCACATGCACCGTCTCAGGGTTCTCATTAACCGCCTATGGTGTAAACTGGGTTCGCCAGCCTCCAGGAAAGGGTCTGGAGTGGCTGGGAATGATATGGGATGATGGAAGCACAGACTATAATTCAGCTCTCAAATCCAGACTGAGCATCAGTAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCAGGTACTACTGTGCCAGAGAAGGGGACGTAGCCTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA SEQ ID NO. 106GACATTGTGATGACACAGTCTCCCTCCTCCCTGGCTGTGTCAGTAGGACAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGCCTTTTAAATGGTAGCAATCAAAAGAACTACTTGGCCTGGTACCAGCAGAAACCAGGACAGTCTCCTAAACTTCTGGTATACTTTGCATCCACTAGGGATTCTGGGGTCCCTGATCGCTTCATAGGCAGTGGATCTGGGACAGATTTCACTCTTACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGATTACTTCTGTCTGCAACATTTTGGCACTCCTCCGACGTTCGGTGGAGGCACCAAACTGGAAATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGTTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTSEQ ID NO. 107CTCCAGCCAGGGGCTGAGGTCCCGGTGGTGTGGGCCCAGGAGGGGGCTCCTGCCCAGCTCCCCTGCAGCCCCACAATCCCCCTCCAGGATCTCAGCCTTCTGCGAAGAGCAGGGGTCACTTGGCAGCATCAGCCAGACAGTGGCCCGCCCGCTGCCGCCCCCGGCCATCCCCTGGCCCCCGGCCCTCACCCGGCGGCGCCCTCCTCCTGGGGGCCCAGGCCCCGCCGCTACACGGTGCTGAGCGTGGGTCCCGGAGGCCTGCGCAGCGGGAGGCTGCCCCTGCAGCCCCGCGTCCAGCTGGATGAGCGCGGCCGGCAGCGCGGGGACTTCTCGCTATGGCTGCGCCCAGCCCGGCGCGCGGACGCCGGCGAGTACCGCGCCGCGGTGCACCTCAGGGACCGCGCCCTCTCCTGCCGCCTCCGTCTGCGCCTGGGCCAGGCCTCGATGACTGCCAGCCCCCCAGGATCTCTCAGAGCCTCCGACTGGGTCATTTTGAACTGCTCCTTCAGCCGCCCTGACCGCCCAGCCTCTGTGCATTGGTTCCGGAACCGGGGCCAGGGCCGAGTCCCTGTCCGGGAGTCCCCCCATCACCACTTAGCGGAAAGCTTCCTCTTCCTGCCCCAAGTCAGCCCCATGGACTCTGGGCCCTGGGGCTGCATCCTCACCTACAGAGATGGCTTCAACGTCTCCATCATGTATAACCTCACTGTTCTGGGTCTGGAGCCCCCAACTCCCTTGACAGTGTACGCTGGAGCAGGTTCCAGGGTGGGGCTGCCCTGCCGCCTGCCTGCTGGTGTGGGGACCCGGTCTTTCCTCACTGCCAAGTGGACTCCTCCTGGGGGAGGCCCTGACCTCCTGGTGACTGGAGACAATGGCGACTTTACCCTTCGACTAGAGGATGTGAGCCAGGCCCAGGCTGGGACCTACACCTGCCATATCCATCTGCAGGAACAGCAGCTCAATGCCACTGTCACATTGGCAATCATCACAGTGACTCCCAAATCCTTTGGGTCACCTGGATCCCTGGGGAAGCTGCTTTGTGAGGTGACTCCAGTATCTGGACAAGAACGCTTTGTGTGGAGCTCTCTGGACACCCCATCCCAGAGGAGTTTCTCAGGACCTTGGCTGGAGGCACAGGAGGCCCAGCTCCTTTCCCAGCCTTGGCAATGCCAGCTGTACCAGGGGGAGAGGCTTCTTGGAGCAGCAGTGTACTTCACAGAGCTGTCTAGCCCACACCACCATCATCACCATSEQ ID NO. 108CCCCAGCCAGGGGCTGAGATCTCGGTGGTGTGGGCCCAGGAGGGGGCTCCTGCCCAGCTCCCCTGCAGCCCCACAATCCCCCTCCAGGATCTCAGCCTTCTGCGAAGAGCAGGGGTCACTTGGCAGCATCAACCAGACAGTGGCCCGCCCGCTCCCGCCCCCGGCCACCCCCCGGCCCCCGGCCATCGCCCGGCGGCGCCCTACTCTTGGGGGCCCAGGCCCCGCCGCTACACAGTGCTGAGCGTGGGTCCTGGAGGCCTGCGCAGCGGGAGGCTGCCCCTGCAGCCCCGCGTCCAGCTGGATGAGCGCGGCCGGCAGCGCGGGGACTTCTCGCTGTGGCTGCGCCCAGCCCGGCGCGCGGACGCCGGCGAGTACCGCGCCACGGTGCACCTCAGGGACCGCGCCCTCTCCTGCCGCCTTCGTCTGCGCGTGGGCCAGGCCTCGATGACTGCCAGCCCCCCAGGGTCTCTCAGGACCTCTGACTGGGTCATTTTGAACTGCTCCTTCAGCCGCCCTGACCGCCCAGCCTCTGTGCATTGGTTCCGGAGCCGTGGCCAGGGCCGAGTCCCTGTCCAGGGGTCCCCCCATCACCACTTAGCGGAAAGCTTCCTCTTCCTGCCCCATGTCGGCCCCATGGACTCTGGGCTCTGGGGCTGCATCCTCACCTACAGAGATGGCTTCAATGTCTCCATCATGTATAACCTCACTGTTCTGGGTCTGGAGCCCGCAACTCCCTTGACAGTGTACGCTGGAGCAGGTTCCAGGGTGGAGCTGCCCTGCCGCCTGCCTCCTGCTGTGGGGACCCAGTCTTTCCTTACTGCCAAGTGGGCTCCTCCTGGGGGAGGCCCTGACCTCCTGGTGGCTGGAGACAATGGCGACTTTACCCTTCGACTAGAGGATGTAAGCCAGGCCCAGGCTGGGACCTACATCTGCCATATCCGTCTACAGGGACAGCAGCTCAATGCCACTGTCACATTGGCAATCATCACAGTGACTCCCAAATCCTTTGGGTCACCTGGCTCCCTGGGGAAGCTGCTTTGTGAGGTGACTCCAGCATCTGGACAAGAACACTTTGTGTGGAGCCCCCTGAACACCCCATCCCAGAGGAGTTTCTCAGGACCATGGCTGGAGGCCCAGGAAGCCCAGCTCCTTTCCCAGCCTTGGCAATGCCAGCTGCACCAGGGGGAGAGGCTTCTTGGAGCAGCAGTATACTTCACAGAACTGTCTAGCCCACACCACCATCATCACCATSEQ ID NO. 109CCCCAGCCAGGGGCTGAGATCTCGGTGGTGTGGGCCCAGGAGGGGGCTCCTGCCCAGCTCCCCTGCAGCCCCACAATCCCCCTCCAGGATCTCAGCCTTCTGCGAAGAGCAGGGGTCACTTGGCAGCATCAACCAGACAGTGGCCCGCCCGCTCCCGCCCCCGGCCACCCCCCGGCCCCCGGCCATCGCCCGGCGGCGCCCTACTCTTGGGGGCCCAGGCCCCGCCGCTACACAGTGCTGAGCGTGGGTCCTGGAGGCCTGCGCAGCGGGAGGCTGCCCCTGCAGCCCCGCGTCCAGCTGGATGAGCGCGGCCGGCAGCGCGGGGACTTCTCGCTGTGGCTGCGCCCAGCCCGGCGCGCGGACGCCGGCGAGTACCGCGCCACGGTGCACCTCAGGGACCGCGCCCTCTCCTGCCGCCTTCGTCTGCGCGTGGGCCAGGCCTCGATGACTGCCAGCCCCCCAGGGTCTCTCAGGACCTCTGACTGGGTCATTTTGAACTGCTCCTTCAGCCGCCCTGACCGCCCAGCCTCTGTGCATTGGTTCCGGAGCCGTGGCCAGGGCCAAGTCCCTGTCCAGGAGTCCCCCCATCACCACTTAGCGGAAAGCTTCCTCTTCCTGCCCCATGTCGGCCCCATGGACTCTGGGCTCTGGGGCTGCATCCTCACCTACAGAGATGGCTTCAATGTCTCCATCATGTATAACCTCACTGTTCTGGGTCTGGAGCCCACAACTCCCTTGACAGTGTACGCTGGAGCAGGTTCCAGGGTGGAGCTGCCCTGCCGCCTGCCTCCTGCTGTGGGGACCCAGTCTTTCCTTACTGCCAAGTGGGCTCCTCCTGGGGGAGGCCCTGACCTCCTGGTGGTTGGAGACAATGGCAACTTTACCCTTCGACTAGAGGATGTAAGCCAGGCCCAGGCTGGGACCTACATCTGCCATATCCGTCTACAGGGACAGCAGCTCAATGCCACTGTCACATTGGCAATCATCACAGTGACTCCCAAATCCTTTGGGTCACCTGGCTCCCTGGGGAAGCTGCTTTGTGAGGTGACTCCAGCATCTGGACAAGAACGCTTTGTGTGGAGCCCCCTGAACACCCCATCCCAGAGGAGTTTCTCAGGACCGTGGCTGGAGGCCCAGGAAGCCCAGCTCCTTTCCCAGCCTTGGCAATGCCAGCTGCACCAGGGGGAGAGGCTTCTTGGAGCAGCAGTATACTTCACAGAACTGTCTAGCCCACACCACCATCATCACCATSEQ ID NO. 110 ATGGGCTGGAGCTGCATCATCCTGTTCCTGGTGGCCACCGCTACCGGAGTGCACAGCSEQ ID NO. 111ATGGAATCACAGACCCAGGTCCTCATGTTTCTTCTGCTCTGGGTATCTGGTGCCTGTGCASEQ ID NO. 112 ATGCCGCTGC TGCTACTGCT GCCCCTGCTG TGGGCAGGGG CGCTAGCT

1. An antigen binding protein which is capable of binding Lymphocyte Activation Gene 3 (LAG-3) and which comprises CDRL1, CDRL2 and CDRL3 from SEQ ID No.
 5. 2. An antigen binding protein according to claim 1, which comprises CDRL1 of SEQ ID NO.
 1. 3. An antigen binding protein according to claim 1 or 2, which comprises CDRL2 of SEQ ID NO. 2 and/or CDRL3 of SEQ ID NO.
 3. 4. An antigen binding protein according to any of claims 1 to 3, which comprises one or more of CDRH1, CDRH2 and CDRH3 from SEQ ID NO.
 10. 5. An antigen binding protein according to any of claims 1 to 4, which comprises one or more CDRs selected from the group comprising CDRH1 of SEQ ID NO. 6, CDRH2 of SEQ ID NO. 7 and CDRH3 of SEQ ID NO.
 8. 6. An antigen binding protein according to any of claims 1 to 5, which comprises the following CDRs: CDRL1: SEQ ID NO. 1, CDRL2: SEQ ID NO. 2, CDRL3: SEQ ID NO. 3, CDRH1: SEQ ID NO. 6, CDRH2: SEQ ID NO. 7, and CDRH3: SEQ ID NO.
 8. 7. An antigen binding protein according to any of claims 1 to 6, which comprises a) the variable light chain (VL) of SEQ ID NO. 4 and b) the variable heavy chain (VH) of SEQ ID No.
 9. 8. An antigen binding protein according to any of claims 1 to 7, which is capable of binding recombinant human sLAG-3 of SEQ ID NO. 51 with a KD of less than 1 nM when determined by Biacore™ surface Plasmon resonance analysis.
 9. An antigen binding protein which is capable of binding recombinant human sLAG-3 of SEQ ID NO. 51 with a KD of less than 1 nM when determined by Biacore™ surface Plasmon resonance analysis.
 10. An antigen binding protein according to any of claims 1 to 9, wherein the antigen binding protein is capable of depleting LAG-3+ activated human T cells.
 11. An antigen binding protein according to claim 10, wherein the antigen binding protein is capable of causing greater than 40% depletion of antigen specific CD4 and/or CD8 LAG-3⁺ human T cells by ADCC in an in-vitro assay using primary human T cells.
 12. An antigen binding protein according to any of claims 1 to 11, wherein the antigen binding protein is a humanised antibody.
 13. A humanised antibody according to claim 12, wherein the humanised antibody constant region is IgG1.
 14. A humanised antibody according to claim 13, which comprises a) a light chain sequence with at least 97% identity to SEQ ID NO. 5, and b) a heavy chain sequence with at least 97% identity to SEQ ID NO.
 10. 15. A humanised antibody according to claim 14, which comprises a) a light chain sequence of SEQ ID NO. 5, and b) a heavy chain sequence of SEQ ID NO.
 10. 16. A humanised antibody according to any of claims 13 to 15, which is non-fucosylated.
 17. An isolated nucleic acid molecule which encodes an antigen binding protein according to any of claims 1 to 12 or an antibody according to any of claims 13 to
 15. 18. An expression vector comprising a nucleic acid molecule according to claim
 17. 19. A host cell comprising an expression vector according to claim
 18. 20. An antigen binding protein as produced by the host cell of claim
 19. 21. A method of producing an antigen binding protein according to any of claims 1 to 12 or an antibody according to any of claims 13 to 15, comprising a) culturing a host cell according to claim 19 and b) isolating the antigen binding protein or antibody.
 22. A method of producing an antibody according to claim 21, comprising a) culturing a host cell according to claim 19, wherein the FUT8 gene encoding alpha-1,6-fucosyltransferase has been inactivated in the recombinant host cell, and b) isolating the antibody.
 23. A pharmaceutical composition comprising a) an antigen binding protein according to any of claims 1 to 12 or an antibody according to any of claims 13 to 16, and b) a pharmaceutically acceptable carrier.
 24. A use of an antigen binding construct as defined in any of claims 1 to 12 or an antibody as defined in any of claims 13 to 16, in therapy.
 25. A use of an antigen binding construct as defined in any of claims 1 to 12 or an antibody as defined in any of claims 13 to 16, in the treatment of diseases associated with the involvement of pathogenic T cells.
 26. A use of an antigen binding construct according to claim 25, wherein the disease is an auto-immune disease or cancer.
 27. A use according to claim 26, wherein the autoimmune disease is selected from the group consisting of psoriasis, Crohn's disease, rheumatoid arthritis, primary biliary cirrhosis, SLE, Sjogren's syndrome, multiple sclerosis, ulcerative colitis and autoimmune hepatitis.
 28. A method of treatment of a human or animal subject comprising administering an antigen binding construct as defined in any of claims 1 to 12 or an antibody as defined in any of claims 13 to
 16. 29. A method of treatment of a disease associated with the involvement of pathogenic T cells in a human or animal subject comprising administering an antigen binding construct as defined in any of claims 1 to 12 or an antibody as defined in any of claims 13 to
 16. 30. A method of treatment according to claim 29, wherein the disease is an autoimmune disease or cancer.
 31. A method of treatment according to claim 30, wherein the autoimmune disease is selected from the group consisting of psoriasis, Crohn's disease, rheumatoid arthritis, primary biliary cirrhosis, SLE, Sjogren's syndrome, multiple sclerosis, ulcerative colitis and autoimmune hepatitis. 